diff --git a/src/contents/results.tsx b/src/contents/results.tsx
index 5cf07760730d949ae8d7e3704a627a7eaecd008c..64d4014626a0dee5e2d92872a9aad3b03664a413 100644
--- a/src/contents/results.tsx
+++ b/src/contents/results.tsx
@@ -21,20 +21,63 @@ export function Results() {
<Section title="Experimental Design" id="ExpDes">
<Subesction title="Proof of Concept" id="Results1">
<h4>acc. to David Liu (Anzalone et al. 2019)</h4>
- <H4 text="Goals"/>
- <p>text</p>
- <H4 text="Workflow"/>
- <p>text</p>
+ <H4 text="Proof-of-concept 1"/>
+ <H5 text="Workflow"/>
+ <p>The prepared pDAS12124-preedited plasmid serves as a positive control to validate the success of the experiment. A technical control with the pZMB938 plasmid confirms successful transfection of the cells. In the main part of the experiment, pDAS12489-2in1 and pCMV-PE2 are co-transfected. Successful transfection is visualised by GFP signals.</p>
<H4 text="Conclusion"/>
- <p>text</p>
+ <p>The microscopy data validates our proof of concept. Compared to our internal positive control, pDAS12124-preedited (see Figure X), less cells co-transfected with pDAS12489 and pCMV-PE2 (see Figure X) showed fluorescence. Contrary to our expectations, the technical transfection control with pZMB938 showed lower transfection efficiency. All negative controls showed no fluorescence.</p>
+ <H4 text="Proof-of-concept 2"/>
+ <H5 text="Workflow"/>
+ <p>To optimise transfection efficiency, different dilutions and concentrations of DNA were used to find the best transfection conditions.</p>
+ <H4 text="Conclusion"/>
+ <p>All 4 different transfection conditions were done with pZMB938 and showed good results, but best result were done when lipofectamine 2000 was diluted 1:10 and 1000 ng DNA was transfected.</p>
+ <H4 text="Proof-of-concept 3"/>
+ <H5 text="Workflow"/>
+ <p>Transfection with Lipofectamine 3000 was performed because of the probably better performance and transfection rate. The prepared pDAS12124-preedited plasmid serves as a positive control to validate the success of the experiment. A technical control with the pZMB938 plasmid confirmed successful transfection of the cells as before. In the main part of the experiment, pDAS12489-2in1 and pCMV-PE2 were co-transfected. Successful transfection and prime editing was detected by GFP signals.</p>
+ <H4 text="Conclusion"/>
+ <p>Internal control and technical control showed higher transfection efficiency then in previous experiments, therefore transfection with lipofectamine 3000 seems to be more efficient than transfection with lipofectamine 2000. The fluorescence of pDAS12189+pCMV-PE2 was still quite low. All negative controls are showed no fluorescence.</p>
+ <H4 text="Proof-of-concept 4"/>
+ <H5 text="Workflow"/>
+ <p>Again a preliminary test with the technical positive control was conducted potentially optimize our transfection protocol and to train the handling.</p>
+ <H4 text="Conclusion"/>
+ <p>The results of the test were not as good as expected. Nearly no transfection efficiency was visible. This could be due to too old hek293 cells</p>
+ <H4 text="Proof-of-concept 5"/>
+ <H5 text="Workflow"/>
+ <p>HEK cells were thawed and another prelimary test was conducted. In this test two different transfection agents were used (Lipofectamine 3000 & CaCl2) to check which one is better suited for our experiments. The literature uses lipofectamine 3000 but CaCl2 transfection is much cheaper.</p>
+ <H4 text="Conclusion"/>
+ <p>Both transfections are working out well but the efficiency of the lipofectamine transfection was much higher.</p>
+ <H4 text="Proof-of-concept 6"/>
+ <H5 text="Workflow"/>
+ <p>One last time the transfection of pDAS12189+pCMV-PE2 was conducted. Although our proof-of-concept already showed successful editing the first time, we repeated the experiment to get better transfection efficiency.</p>
+ <H4 text="Conclusion"/>
+ <p>The transfection efficiency was much better. Our proof-of-concept was working correctly. The reporter system pDAS12189 only led to production of a fluorescent signal when co transfected with a prime editing complex as pCMV-PE2.</p>
</Subesction>
<Subesction title="Mechanism" id="Results2">
- <H4 text="Goals"/>
- <p>text</p>
- <H4 text="Workflow"/>
- <p>text</p>
- <H4 text="Conclusion"/>
- <p>text</p>
+ <H4 text="pegRNA 1"/>
+ <H5 text="Workflow"/>
+ <p>With this experiment we wanted to compare the efficiency of pegRNAs with and without silent edits.</p>
+ <H5 text="Conclusion"/>
+ <p>The FACS analysis shows that pegRNA without silent edits (pegRNA1) had a 2.05 times higher transfection efficiency than pegRNA with silent edits (pegRNA2).</p>
+ <H4 text="pegRNA 2"/>
+ <H5 text="Workflow"/>
+ <p>Cotransfection of pPEAR_CFTR and PE2 and also 1 of the 14 pegRNAs to compare the transfection efficiency of all of our designed pegRNAs.</p>
+ <H5 text="Conclusion"/>
+ <p>The pegRNAs lead to differing amounts of cells showing fluorescence, which, assuming comparable transfection efficiencies, indicates varying prime editing efficiency. The pegRNA7 showed the highest transfection efficiency (see Figure X).</p>
+ <H4 text="pegRNA 3"/>
+ <H5 text="Workflow"/>
+ <p>We tried to transfect CFBE41o- cells with pDAS12124-preedited, our internal positive control, to check if a transfection of this cell line is possible. Furthermore we tried to co transfect the CFBE41o- with pPEAR_CFTR, PE6c and pegRNA4.</p>
+ <H5 text="Conclusion"/>
+ <p>Transfection of CFBE41o- with pDAS12124-preedited was successful (see Figure X). After 24 hours a successful co transfection of pPEAR_CFTR with PE6c and pegRNA4 was visible, although the transfection efficiency was really bad (see Figure X).</p>
+ <H4 text="pegRNA 4"/>
+ <H5 text="Workflow"/>
+ <p>pCMV-PE2 was co transfected with pDAS12489 and pCMV-PE_CO-Mini was co transfected with pDAS12489 in HEK293 cell line.</p>
+ <H5 text="Conclusion"/>
+ <p>The FACS results show that transfection with pCMV-PE2 as the prime editing complex had editing efficiency of 52.90% when normalized on pDAS12124-preedited. When pCMV-PE_CO-Mini was used as a prime editing complex it had a transfection efficiency of 2.54% (see Figure X).</p>
+ <H4 text="pegRNA 5"/>
+ <H5 text="Workflow"/>
+ <p>We compared the 3 different Prime Editing complexes (pCMV-PE2, pCMV-PE2_CO-Mini & pCMV-PE6c) to check which one has the best transfection efficiency.</p>
+ <H5 text="Conclusion"/>
+ <p>The FACS measurement shows the fluorescence rate cells co-transfected with pDAS12489 and pCMV-PE6c as a prime editing complex. The editing efficiency off PE6c was by far the highest (81.88%) (see Figure X). The efficiency was 1.55 higher than the efficiency when pCMV-PE2 was used as prime editing complex (see Figure X).</p>
</Subesction >
<Subesction title="Delivery" id="Results3">
<H4 text="RNA Synthesis"/>
diff --git a/src/contents/safety.tsx b/src/contents/safety.tsx
index 3de4b307a944c723bbf3a17063886f2241c26cdc..8a20472cf71ce83864d5302e71aa7fed7a1949f9 100644
--- a/src/contents/safety.tsx
+++ b/src/contents/safety.tsx
@@ -21,149 +21,7 @@ export const Safety: React.FC = () =>{
As part of our project <PreCyse/> to develop a prime-editing complex to correct the F508del mutation in cystic fibrosis, we place great emphasis on safety at all stages of research. Our final construct will be tested in <a onClick={() => goToPageAndScroll ('Cell Culture3H', '/materials-methods')}> primary cultures of nasal epithelial cells </a> obtained from nasal swabs, isolated from both patients and healthy individuals. To guarantee safety and ensure the highest level of precision and reliability of our results, we have introduced a series of carefully planned checkpoints during the experiments. These milestones allow for continuous monitoring, timely adjustments and validation at each critical stage. This ensures that potential issues are identified and addressed immediately, minimizing risk and improving the overall quality of the experimental results.
</p>
</Section>
- <Section title="Check-Ins" id="Check-Ins">
- <div>
- <p>
- iGEM places great emphasis on biosafety, ensuring that all projects adhere to strict safety standards. One of these measures is the iGEM White List, which includes organisms and parts that are pre-approved for use based on their safety profile. Any components or organisms not covered by this White List must be submitted as 'Check-Ins' to the iGEM Safety Committee for approval. Check-Ins are formal safety evaluations that allow the committee to assess the potential risks and ensure proper containment and handling procedures are in place. Although we used some parts and organisms that were not included on the White List, these were assessed as critical for our project and submitted as Check-Ins to the iGEM Safety Committee. Furthermore, we were in active exchange with the committee throughout the process. The Check-Ins provide a clear picture of the biosafety aspects of our project, reflecting our commitment to safety and compliance with iGEM standards.
- </p>
- <p>
- We adhere to good laboratory practices by ensuring proper handling of materials, effective emergency procedures, and correct waste disposal methods. This commitment guarantees a safe and compliant research environment. Our project, which involved a wide range of techniques was conducted in strict compliance with safety regulations. All experiments were carried out in Prof. Dr. Kristian Müller’s laboratory at Bielefeld University, following BSL-1 standard operating procedures. Properly equipped facilities are crucial to prevent contamination, exposure, or accidental release of modified organisms, ensuring the highest level of safety in our laboratories.
- Before commencing laboratory work, all participants were required to attend a mandatory safety briefing. In compliance with German regulations, each team member's participation had to be confirmed with a personal signature. The briefing, conducted by Prof. Dr. Kristian Müller must be renewed annually in accordance with §12 ArbSchG. It covered the following areas:
- </p>
- <ul>
- <li>General laboratory safety</li>
- <li>Regulations regarding hazardous and toxic substances</li>
- <li>Regulations concerning biological materials</li>
- <li>Regulations on genetic engineering</li>
- </ul>
- <p>
- In addition to the general safety briefing, specific instructions for the safe operation of each device were provided. The Safety and Security Officer within the laboratory highlighted the potential hazards and necessary precautionary measures. We have focused on planning our laboratory activities to minimize risk for safer practices. This ensures not only the safe and proper use of equipment but also the generation of reliable data. To meet all safety requirements, additional safety protocols have been put in place for all targeted areas of the laboratory equipment.
- </p>
- <H4 text="Laboratory and safety practices"></H4>
- <p>
- As part of our project to develop a prime-editing complex to correct the F508del mutation in cystic fibrosis, we place great emphasis on safety at all stages of research. Our final construct will be tested in <a onClick={() => goToPageAndScroll ('Cell Culture3H', '/materials-methods')}> primary cultures of epithelial cells </a> obtained from nasal swabs, isolated from both patients and healthy individuals. To guarantee safety and ensure the highest level of precision and reliability of our results, we have introduced a series of carefully planned checkpoints during the experiments. These milestones allow for continuous monitoring, timely adjustments and validation at each critical stage. This ensures that potential issues are identified and addressed immediately, minimizing risk and improving the overall quality of the experimental results. [link zu den Experimenten] . iGEM places great emphasis on biosafety, ensuring that all projects adhere to strict safety standards. One of these measures is the iGEM White List, which includes organisms and parts that are pre-approved for use based on their safety profile. Any components or organisms not covered by this White List must be submitted as 'Check-Ins' to the iGEM Safety Committee for approval. Check-Ins are formal safety evaluations that allow the committee to assess the potential risks and ensure proper containment and handling procedures are in place. Although we used some parts and organisms that were not included on the White List, these were assessed as critical for our project and submitted as Check-Ins to the iGEM Safety Committee. Furthermore, we were in active exchange with the committee throughout the process. The Check-ins provide a clear picture of the biosafety aspects of our project, reflecting our commitment to safety and compliance with iGEM standards.
- The main safety measures we have implemented include:
- </p>
- <p>
- <strong>Compliance with S1 conditions:</strong> Working in S1 laboratories ensures that only organisms in the lowest risk group are used, minimizing the risk to humans and the environment.
- </p>
- <p>
- <strong>Sterile working practices:</strong> To avoid contamination, we have implemented strict hygiene measures, including the disinfection of work surfaces and the correct disposal of biological waste.
- </p>
- <p>
- <strong>Controlled access:</strong> Access to laboratories was strictly regulated to ensure that only trained personnel worked with the genetically modified organisms and cell lines.
- </p>
- <p>
- <strong>Documentation:</strong> All work steps, materials used and cell lines were carefully documented to ensure traceability and safety.
- </p>
- <p>
- <strong>Safe handling of cell lines:</strong> The cell lines used for experiments were handled in accordance with the applicable safety regulations. This included regular checks for contamination and the safe storage and disposal of cell cultures.
- </p>
- <Collapsible id="Checkpek" open={false} title="Check-in for the Prime-Editing Komplex ">
- <p>
- <strong>Reverse transcriptase:</strong> Reverse transcriptase plays a central role in prime editing by specifically inserting the correction as DNA at the inserted nick using an RNA template provided by pegRNA. The correction of the complementary DNA strand then takes place via the natural cell repair mechanisms. This ensures an exact correction of the target sequence. We checked the reverse transcriptase to ensure it could perform precise genome editing without introducing unintended mutations. This was important to minimize the risk of off-target effects that could lead to unexpected or harmful consequences.
- </p>
- <p>
- <strong>pegRNA (Prime Editing Guide RNA):</strong> The pegRNA is a multifunctional RNA molecule that fulfils two essential tasks. Firstly, it serves as a standard guide RNA (gRNA) that binds specifically to the target DNA and thus marks the site of editing. Secondly, it contains an RNA template that encodes the desired DNA modification. This enables the precise integration of the genetic modifications at the target site. We evaluated pegRNA for its ability to specifically target and modified the intended DNA sequence. Ensuring its specificity was crucial to avoid the potential disruption of other genes.
- </p>
- <p>
- <strong>Nickase nCas9, CasX, Fanzor (SpuFz1):</strong> These modified nucleases are designed to cut only one strand of DNA. This leads to controlled and precise editing of the genome, as cutting only one strand minimizes the risk of unwanted double-strand breaks. CasX and Fanzor offer smaller alternatives to Cas9, which is particularly advantageous for use in cells or organisms where space and efficiency requirements in terms of the transport system are an issue. Fanzor, being a newly introduced endonuclease, was particularly scrutinized in our project to ensure its safety and effectiveness in different cellular contexts.
-
- This prime-editing complex thus represents a precise and efficient method for gene editing. By combining these components, genetic modifications can be performed with minimal side effects
- </p>
- </Collapsible>
-
- <Collapsible id="Checkcloning" open={false} title="Check-in for Cloning">
- <p>
- For our cloning experiments and the development of our prime editing complexes, we have amplified various plasmids in <i>E. coli</i> K-12 strains (DH5α,10-Beta). When working with microbial strains such as <i>E. coli</i> K-12 strains, it's important to consider potential risks associated with their use, even though they are generally regarded as safe in laboratory settings. All experiments were performed under strict S1 conditions, following all relevant safety protocols. Below you will find an overview of the <i>E. coli</i> K-12 strains for our cloning experiments, submitted by us as a check-In and the specific safety measures:
- </p>
- <p>
- <strong><i>E. coli K-12</i> strains (DH5α, 10-Beta):</strong> Although these strains are non-pathogenic and have been modified to minimize the risk of spreading antibiotic resistance, there remains a low risk of horizontal gene transfer, where genetic material could be transferred to other microorganisms, potentially leading to the spread of resistance genes or other traits. If accidentally released into the environment, <i>E. coli</i> K-12 strains could potentially interact with native microbial communities. While they are typically outcompeted in natural environments, there's a remote possibility of ecological disruption, particularly in microenvironments where they could find a niche.While these strains are non-virulent, they still pose a minimal risk to humans, particularly immunocompromised individuals, through accidental ingestion or inhalation in a laboratory setting.
- </p>
- <p>
- We submitted the yeast strain <i>Pichia pastoris</i> (SMD1163) for the protein expression of Fanzor.
- </p>
- <p>
- <strong><i>Pichia pastoris</i> (SMD1163):</strong> <i>Pichia pastoris</i> (SMD1163) is a widely used yeast strain for the expression of recombinant proteins. It is characterized by a methanol-inducible expression system (AOX1 promoter) and high cell growth rates, which makes it ideal for industrial applications. The strain can be easily genetically manipulated and can perform post-translational modifications, which supports correct protein production.
- When working with <i>Pichia pastoris</i> (SMD1163), various safety-relevant aspects must be observed. Although the organism is considered non-pathogenic and biologically safe (S1), skin contact and aerosol formation should be avoided to minimize the risk of infection or allergic reactions. When using genetically modified strains, it is important to follow the relevant GMO guidelines to prevent uncontrolled release. In addition, handling chemicals such as methanol requires special precautions as they are toxic and highly flammable. The disposal of cell cultures and waste must also be carried out in accordance with biosafety regulations, especially in the case of genetically modified organisms.
- </p>
- </Collapsible>
-
- <Collapsible id="CheckcellLines" open={false} title="Check-in for Testing in cell lines">
- <p>
- In our project, we paid attention to safety at every step, especially when working with specific <a onClick={() => goToPageAndScroll ('cell-culture', '/materials-methods')}> cell lines </a> . All experiments were performed under strict S1 conditions, following all relevant safety protocols. Given the sensitivity of the human cell lines we used, we placed great emphasis on controlled and well-designed workflows. All transfections were performed in our own transfection laboratory to ensure a high level of safety and compliance. Below you will find an overview of the cell lines submitted by us as a checkin and the specific safety measures:
- </p>
- <p>
- <strong>HEK293 cell line: </strong>HEK 293 (Human Embryonic Kidney 293) cells are an immortal cell line originally derived from the kidney cells of a human embryo. They are characterized by their fast division rate and high transfection efficiency, which makes them a popular model in biomedical research. For our studies, the basic HEK293 cells were provided to us by the Cellular and Molecular Biotechnology Group at Bielefeld University, headed by Prof. Dr. Kristian Müller. Prof. Dr. Müller is also one of the Principal Investigators of our team. We use this cell line in our proof-of-concept studies and for testing the Prime Editing Guide pegRNA (pegRNA) to evaluate the efficiency and functionality of our constructs.
- </p>
- <p>
- <strong>HEK293T-3HA-CFTR cell line: </strong>The HEK293T-3HA-CFTR cell line is based on HEK293T cells expressing an additional tsA1609 allele of the SV40 large T antigen. This allele enables the replication of vectors containing the SV40 origin of replication. In addition to the native CFTR gene, which is not expressed in HEK cells, the HEK293T-3HA-CFTR cell line carries another copy of the CFTR gene embedded in an expression cassette. This cassette contains a CMV promoter, which is derived from the human cytomegalovirus and is frequently used for the overexpression of genes in human cells. In addition, the cassette contains a puromycin resistance gene that is co-expressed with CFTR, allowing continuous selection of CFTR-expressing cells.
- </p>
- <p>
- <strong>HEK293T-3HA-F508del-CFTR cell line:</strong> The HEK293T-3HA-F508del-CFTR cell line is a modified HEK293T cell line that carries the F508del mutation in the CFTR gene, which is responsible for the most common mutation in cystic fibrosis. This mutation leads to a defective CFTR protein that impairs the normal function of the chloride channel. The cell line is therefore ideal for studying the effects of this mutation and for evaluating potential therapies for cystic fibrosis.
- </p>
- <p>
- <strong>CFBE41o- cell line:</strong> The CFBE41o- cell line, derived from the bronchial epithelial cells of a cystic fibrosis patient, is homozygous for the F508del-CFTR mutation and was essential for our cystic fibrosis research. A reduced CFTR expression level is present. The cell line carries the CFTR defect and can therefore represent a patient with CF. The cell line is used to test our mechanism. These cells were immortalized with a replication-defective plasmid that retains their physiological properties.
- </p>
- <p>
- When working with the HEK293T and CFBE41o- cell lines, it’s important to consider the minimal risks associated with their use. While not harmful on their own, the genetic modifications in HEK293T cells require careful handling to prevent accidental release or exposure. These cells, engineered to overexpress CFTR, including the F508del mutation, necessitate strict safety measures like regular monitoring and proper waste disposal to comply with S1 laboratory standards. Similarly, CFBE41o- cells, due to their genetic modifications and disease relevance, require careful handling to avoid cross-contamination and ensure biosafety.
- </p>
- <p>
- <strong>Human nasal epithelial cells (hNECs):</strong> Human nasal epithelial cells (hNECs) were harvested using a nasal brush, a minimally invasive procedure, and cultured in air-liquid interface (ALI) cultures to model the airway epithelium. Human nasal epithelial cells (hNECs) were obtained using a nasal brush, a minimally invasive technique, and then cultured in air-liquid interface (ALI) cultures to model the airway epithelium. Using these primary cultures, derived from donors with airway diseases such as cystic fibrosis, we were able to simulate the in vivo conditions of such diseases.
- Due to the sensitive nature of these primary human cells, we performed all experiments with hNECs in our S2 laboratory, where increased safety precautions were taken. This included strict safety controls, safe handling of samples and proper disposal of materials after testing. In particular, the hNECs underwent HHH (Triple H: HIV, HCV and HBV) testing to ensure that no contamination occurred during sample collection or experimentation. These tests included sterility testing, viability assessments and contamination testing to ensure the safety and integrity of both the samples and the laboratory environment. After a negative HHH test, the primary cultures can be treated as S1. In addition, the nasal epithelial cells were handled with the utmost care during collection, ensuring that all procedures were performed under sterile conditions to avoid any risk of contaminationFor this purpose, the intensive examination of ethical questions was fundamental and a constant companion of our project. The numerous results from the interviews in the areas of: Ethics, storage and training in the handling of samples have been summarized in a guideline for patient consent for Germany and are intended to provide iGEM teams with the scope, critical examination and observance of iGEM rules, international and national guidelines. 
- </p>
- </Collapsible>
-
- <Collapsible id="CheckDelivery" open={false} title="Check-in for Delivery">
- <p>
- Our finished construct is designed to be delivered into the lung via an inhaler using lipid nanoparticles (LNPs). To be more spezific a selective organ-targeting (SORT)- LNPs were developed to deliver mRNA specifically to the lung, with special measures taken to increase biocompatibility and safety. Since the LNP composition is very specific and also differs from other formulas, we submitted the LNP as a checkin:
- </p>
- <p>
- <strong>LNP:</strong> These LNPs are then taken up by epithelial cells through endocytosis, releasing the construct into the cytosol. We carefully evaluated the potential risks, including unintended immune responses and the need for precise dosing to minimize side effects. In addition, we have conducted an in-depth analysis of the dual-use potential of our technology. Dual-use refers to the possibility that scientific advances can be used for both civilian and military purposes. Therefore, we have implemented strict safety protocols and ethical guidelines to ensure that our technology is used exclusively for peaceful and therapeutic applications.
- </p>
- </Collapsible>
- </div>
- </Section>
- <Section title="Our Lab" id="Our Lab">
- <p>
- As part of our laboratory activities for our <PreCyse/> project, we worked in various laboratories. For general lab work and cloning experiments, you can find some pictures of our laboratories below:
- </p>
- <H4 text="Our Cloning Lab"></H4>
- <p>
- Our Cloning-laboratory is divided into different work areas to ensure that the experiments run smoothly and efficiently. These include the gel station, the PCR station, the transformation section and the measurement area. Each area is specially equipped for the respective method, and the corresponding experiments were carried out exclusively in the designated stations. In this way, we ensure that our work is carried out under optimal conditions and with the greatest possible precision.
- <div className="figure-wrapper">
- <figure>
- <img src="https://static.igem.wiki/teams/5247/photos/biosafety/kollage/new/img-2041.jpeg" style={{height: "10%", width:"auto"}}/>
- <figcaption> <b>Figure 1</b> Photo-gallery of laboratory. A: Key lock. B: Key-locked door. C: Alarm plan. D: Emergeny button for electriotion stop. E: Emergency telephone. F: First aid kit, cardiac defibrillaton and emergency exit and fire alarm plan. G: Wash bin with emergency eye wash. H: Emergency shower. I: Lockable cabinets for chemical storage. </figcaption>
- </figure>
- </div>
- <div className="figure-wrapper">
- <figure>
- <img src="https://static.igem.wiki/teams/5247/photos/biosafety/kollage/new/img-2037.jpeg" style={{height: "10%", width: "auto"}}/>
- <figcaption> <b>Figure 2</b> Photo-gallery of S1 laboratory. A: Autoclave. B: Refrigerator with chemicals. C: Weighing room with chemical storage. D: Clean bench work space with vortex, pipettes, heat block and bench top centrifuge. E: pH electrode in fume hood. F: Ice machine. G: Fire distinguisher and S1 waste. H: Fume hood with liquid waste.</figcaption>
- </figure>
- </div>
- </p>
- <H4 text="Our Cell Culture Lab "></H4>
- <p>
- In our cell culture laboratory, we work under sterile conditions to ensure optimal growth conditions for human cell lines. Among other things, we carry out transfections in order to introduce genetic material into cells and investigate their behavior. Strict protocols and state-of-the-art technology ensure the precision and reproducibility of our experiments.
- </p>
- <div className="figure-wrapper">
- <figure>
- <img src="https://static.igem.wiki/teams/5247/photos/biosafety/kollage/new/img-2040.jpeg" style={{height: "10%", width: "auto"}}/>
- <figcaption> <b>Figure 3</b> Photo-gallery of laboratory and chemical storage. A: Safety cabinets. B: Incubator. C: Safety cabinet.</figcaption>
- </figure>
- </div>
- <p>
- In our S2 laboratory, the harvested nasal epithelial cells that serve as primary cultures undergo a comprehensive HHH test to ensure their safety and suitability for further experiments. This test is crucial to ensure that we can subsequently work safely with these cells in the S1 range without the risk of contamination or unwanted release of biological material.
- </p>
- <div className="figure-wrapper">
- <figure>
- <img src="https://static.igem.wiki/teams/5247/photos/biosafety/kollage/new/img-2042.jpeg" style={{height: "10%"}}/>
- <figcaption> <b>Figure 4</b>Photo-gallery of S2 laboratory. A: Door of S2 lab with S2 sign. B: Emergency shower and fire distinguisher. C: Clean bench with centrifuge. D: Incubator. E: Safety cabinet. F: Emergeny telephone. G: S2 lab coat with S2 sign. H: Microscope. I: Autoclave. </figcaption>
- </figure>
- </div>
- </Section>
+
<Section title="Biosafety" id="Biosafety">
<Subesction title="Safety aspects of our PrimeGuide" id="Biosafety1">
<p>
@@ -317,7 +175,6 @@ export const Safety: React.FC = () =>{
This proactive approach helps us address potential concerns, demystify our research, and contribute to a more informed public perception of synthetic biology.
</p>
</Subesction>
- </Section>
<Section title="Bioethics" id="Bioethics">
<div>
<p>
@@ -360,6 +217,150 @@ export const Safety: React.FC = () =>{
<LoremMedium/>
</Subesction>
</Section>
+ <Section title="Check-Ins" id="Check-Ins">
+ <div>
+ <p>
+ iGEM places great emphasis on biosafety, ensuring that all projects adhere to strict safety standards. One of these measures is the iGEM White List, which includes organisms and parts that are pre-approved for use based on their safety profile. Any components or organisms not covered by this White List must be submitted as 'Check-Ins' to the iGEM Safety Committee for approval. Check-Ins are formal safety evaluations that allow the committee to assess the potential risks and ensure proper containment and handling procedures are in place. Although we used some parts and organisms that were not included on the White List, these were assessed as critical for our project and submitted as Check-Ins to the iGEM Safety Committee. Furthermore, we were in active exchange with the committee throughout the process. The Check-Ins provide a clear picture of the biosafety aspects of our project, reflecting our commitment to safety and compliance with iGEM standards.
+ </p>
+ <p>
+ We adhere to good laboratory practices by ensuring proper handling of materials, effective emergency procedures, and correct waste disposal methods. This commitment guarantees a safe and compliant research environment. Our project, which involved a wide range of techniques was conducted in strict compliance with safety regulations. All experiments were carried out in Prof. Dr. Kristian Müller’s laboratory at Bielefeld University, following BSL-1 standard operating procedures. Properly equipped facilities are crucial to prevent contamination, exposure, or accidental release of modified organisms, ensuring the highest level of safety in our laboratories.
+ Before commencing laboratory work, all participants were required to attend a mandatory safety briefing. In compliance with German regulations, each team member's participation had to be confirmed with a personal signature. The briefing, conducted by Prof. Dr. Kristian Müller must be renewed annually in accordance with §12 ArbSchG. It covered the following areas:
+ </p>
+ <ul>
+ <li>General laboratory safety</li>
+ <li>Regulations regarding hazardous and toxic substances</li>
+ <li>Regulations concerning biological materials</li>
+ <li>Regulations on genetic engineering</li>
+ </ul>
+ <p>
+ In addition to the general safety briefing, specific instructions for the safe operation of each device were provided. The Safety and Security Officer within the laboratory highlighted the potential hazards and necessary precautionary measures. We have focused on planning our laboratory activities to minimize risk for safer practices. This ensures not only the safe and proper use of equipment but also the generation of reliable data. To meet all safety requirements, additional safety protocols have been put in place for all targeted areas of the laboratory equipment.
+ </p>
+ <H4 text="Laboratory and safety practices"></H4>
+ <p>
+ As part of our project to develop a prime-editing complex to correct the F508del mutation in cystic fibrosis, we place great emphasis on safety at all stages of research. Our final construct will be tested in <a onClick={() => goToPageAndScroll ('Cell Culture3H', '/materials-methods')}> primary cultures of epithelial cells </a> obtained from nasal swabs, isolated from both patients and healthy individuals. To guarantee safety and ensure the highest level of precision and reliability of our results, we have introduced a series of carefully planned checkpoints during the experiments. These milestones allow for continuous monitoring, timely adjustments and validation at each critical stage. This ensures that potential issues are identified and addressed immediately, minimizing risk and improving the overall quality of the experimental results. [link zu den Experimenten] . iGEM places great emphasis on biosafety, ensuring that all projects adhere to strict safety standards. One of these measures is the iGEM White List, which includes organisms and parts that are pre-approved for use based on their safety profile. Any components or organisms not covered by this White List must be submitted as 'Check-Ins' to the iGEM Safety Committee for approval. Check-Ins are formal safety evaluations that allow the committee to assess the potential risks and ensure proper containment and handling procedures are in place. Although we used some parts and organisms that were not included on the White List, these were assessed as critical for our project and submitted as Check-Ins to the iGEM Safety Committee. Furthermore, we were in active exchange with the committee throughout the process. The Check-ins provide a clear picture of the biosafety aspects of our project, reflecting our commitment to safety and compliance with iGEM standards.
+ The main safety measures we have implemented include:
+ </p>
+ <p>
+ <strong>Compliance with S1 conditions:</strong> Working in S1 laboratories ensures that only organisms in the lowest risk group are used, minimizing the risk to humans and the environment.
+ </p>
+ <p>
+ <strong>Sterile working practices:</strong> To avoid contamination, we have implemented strict hygiene measures, including the disinfection of work surfaces and the correct disposal of biological waste.
+ </p>
+ <p>
+ <strong>Controlled access:</strong> Access to laboratories was strictly regulated to ensure that only trained personnel worked with the genetically modified organisms and cell lines.
+ </p>
+ <p>
+ <strong>Documentation:</strong> All work steps, materials used and cell lines were carefully documented to ensure traceability and safety.
+ </p>
+ <p>
+ <strong>Safe handling of cell lines:</strong> The cell lines used for experiments were handled in accordance with the applicable safety regulations. This included regular checks for contamination and the safe storage and disposal of cell cultures.
+ </p>
+ <Collapsible id="Checkpek" open={false} title="Check-in for the Prime-Editing Komplex ">
+ <p>
+ <strong>Reverse transcriptase:</strong> Reverse transcriptase plays a central role in prime editing by specifically inserting the correction as DNA at the inserted nick using an RNA template provided by pegRNA. The correction of the complementary DNA strand then takes place via the natural cell repair mechanisms. This ensures an exact correction of the target sequence. We checked the reverse transcriptase to ensure it could perform precise genome editing without introducing unintended mutations. This was important to minimize the risk of off-target effects that could lead to unexpected or harmful consequences.
+ </p>
+ <p>
+ <strong>pegRNA (Prime Editing Guide RNA):</strong> The pegRNA is a multifunctional RNA molecule that fulfils two essential tasks. Firstly, it serves as a standard guide RNA (gRNA) that binds specifically to the target DNA and thus marks the site of editing. Secondly, it contains an RNA template that encodes the desired DNA modification. This enables the precise integration of the genetic modifications at the target site. We evaluated pegRNA for its ability to specifically target and modified the intended DNA sequence. Ensuring its specificity was crucial to avoid the potential disruption of other genes.
+ </p>
+ <p>
+ <strong>Nickase nCas9, CasX, Fanzor (SpuFz1):</strong> These modified nucleases are designed to cut only one strand of DNA. This leads to controlled and precise editing of the genome, as cutting only one strand minimizes the risk of unwanted double-strand breaks. CasX and Fanzor offer smaller alternatives to Cas9, which is particularly advantageous for use in cells or organisms where space and efficiency requirements in terms of the transport system are an issue. Fanzor, being a newly introduced endonuclease, was particularly scrutinized in our project to ensure its safety and effectiveness in different cellular contexts.
+
+ This prime-editing complex thus represents a precise and efficient method for gene editing. By combining these components, genetic modifications can be performed with minimal side effects
+ </p>
+ </Collapsible>
+
+ <Collapsible id="Checkcloning" open={false} title="Check-in for Cloning">
+ <p>
+ For our cloning experiments and the development of our prime editing complexes, we have amplified various plasmids in <i>E. coli</i> K-12 strains (DH5α,10-Beta). When working with microbial strains such as <i>E. coli</i> K-12 strains, it's important to consider potential risks associated with their use, even though they are generally regarded as safe in laboratory settings. All experiments were performed under strict S1 conditions, following all relevant safety protocols. Below you will find an overview of the <i>E. coli</i> K-12 strains for our cloning experiments, submitted by us as a check-In and the specific safety measures:
+ </p>
+ <p>
+ <strong><i>E. coli K-12</i> strains (DH5α, 10-Beta):</strong> Although these strains are non-pathogenic and have been modified to minimize the risk of spreading antibiotic resistance, there remains a low risk of horizontal gene transfer, where genetic material could be transferred to other microorganisms, potentially leading to the spread of resistance genes or other traits. If accidentally released into the environment, <i>E. coli</i> K-12 strains could potentially interact with native microbial communities. While they are typically outcompeted in natural environments, there's a remote possibility of ecological disruption, particularly in microenvironments where they could find a niche.While these strains are non-virulent, they still pose a minimal risk to humans, particularly immunocompromised individuals, through accidental ingestion or inhalation in a laboratory setting.
+ </p>
+ <p>
+ We submitted the yeast strain <i>Pichia pastoris</i> (SMD1163) for the protein expression of Fanzor.
+ </p>
+ <p>
+ <strong><i>Pichia pastoris</i> (SMD1163):</strong> <i>Pichia pastoris</i> (SMD1163) is a widely used yeast strain for the expression of recombinant proteins. It is characterized by a methanol-inducible expression system (AOX1 promoter) and high cell growth rates, which makes it ideal for industrial applications. The strain can be easily genetically manipulated and can perform post-translational modifications, which supports correct protein production.
+ When working with <i>Pichia pastoris</i> (SMD1163), various safety-relevant aspects must be observed. Although the organism is considered non-pathogenic and biologically safe (S1), skin contact and aerosol formation should be avoided to minimize the risk of infection or allergic reactions. When using genetically modified strains, it is important to follow the relevant GMO guidelines to prevent uncontrolled release. In addition, handling chemicals such as methanol requires special precautions as they are toxic and highly flammable. The disposal of cell cultures and waste must also be carried out in accordance with biosafety regulations, especially in the case of genetically modified organisms.
+ </p>
+ </Collapsible>
+
+ <Collapsible id="CheckcellLines" open={false} title="Check-in for Testing in cell lines">
+ <p>
+ In our project, we paid attention to safety at every step, especially when working with specific <a onClick={() => goToPageAndScroll ('cell-culture', '/materials-methods')}> cell lines </a> . All experiments were performed under strict S1 conditions, following all relevant safety protocols. Given the sensitivity of the human cell lines we used, we placed great emphasis on controlled and well-designed workflows. All transfections were performed in our own transfection laboratory to ensure a high level of safety and compliance. Below you will find an overview of the cell lines submitted by us as a checkin and the specific safety measures:
+ </p>
+ <p>
+ <strong>HEK293 cell line: </strong>HEK 293 (Human Embryonic Kidney 293) cells are an immortal cell line originally derived from the kidney cells of a human embryo. They are characterized by their fast division rate and high transfection efficiency, which makes them a popular model in biomedical research. For our studies, the basic HEK293 cells were provided to us by the Cellular and Molecular Biotechnology Group at Bielefeld University, headed by Prof. Dr. Kristian Müller. Prof. Dr. Müller is also one of the Principal Investigators of our team. We use this cell line in our proof-of-concept studies and for testing the Prime Editing Guide pegRNA (pegRNA) to evaluate the efficiency and functionality of our constructs.
+ </p>
+ <p>
+ <strong>HEK293T-3HA-CFTR cell line: </strong>The HEK293T-3HA-CFTR cell line is based on HEK293T cells expressing an additional tsA1609 allele of the SV40 large T antigen. This allele enables the replication of vectors containing the SV40 origin of replication. In addition to the native CFTR gene, which is not expressed in HEK cells, the HEK293T-3HA-CFTR cell line carries another copy of the CFTR gene embedded in an expression cassette. This cassette contains a CMV promoter, which is derived from the human cytomegalovirus and is frequently used for the overexpression of genes in human cells. In addition, the cassette contains a puromycin resistance gene that is co-expressed with CFTR, allowing continuous selection of CFTR-expressing cells.
+ </p>
+ <p>
+ <strong>HEK293T-3HA-F508del-CFTR cell line:</strong> The HEK293T-3HA-F508del-CFTR cell line is a modified HEK293T cell line that carries the F508del mutation in the CFTR gene, which is responsible for the most common mutation in cystic fibrosis. This mutation leads to a defective CFTR protein that impairs the normal function of the chloride channel. The cell line is therefore ideal for studying the effects of this mutation and for evaluating potential therapies for cystic fibrosis.
+ </p>
+ <p>
+ <strong>CFBE41o- cell line:</strong> The CFBE41o- cell line, derived from the bronchial epithelial cells of a cystic fibrosis patient, is homozygous for the F508del-CFTR mutation and was essential for our cystic fibrosis research. A reduced CFTR expression level is present. The cell line carries the CFTR defect and can therefore represent a patient with CF. The cell line is used to test our mechanism. These cells were immortalized with a replication-defective plasmid that retains their physiological properties.
+ </p>
+ <p>
+ When working with the HEK293T and CFBE41o- cell lines, it’s important to consider the minimal risks associated with their use. While not harmful on their own, the genetic modifications in HEK293T cells require careful handling to prevent accidental release or exposure. These cells, engineered to overexpress CFTR, including the F508del mutation, necessitate strict safety measures like regular monitoring and proper waste disposal to comply with S1 laboratory standards. Similarly, CFBE41o- cells, due to their genetic modifications and disease relevance, require careful handling to avoid cross-contamination and ensure biosafety.
+ </p>
+ <p>
+ <strong>Human nasal epithelial cells (hNECs):</strong> Human nasal epithelial cells (hNECs) were harvested using a nasal brush, a minimally invasive procedure, and cultured in air-liquid interface (ALI) cultures to model the airway epithelium. Human nasal epithelial cells (hNECs) were obtained using a nasal brush, a minimally invasive technique, and then cultured in air-liquid interface (ALI) cultures to model the airway epithelium. Using these primary cultures, derived from donors with airway diseases such as cystic fibrosis, we were able to simulate the in vivo conditions of such diseases.
+ Due to the sensitive nature of these primary human cells, we performed all experiments with hNECs in our S2 laboratory, where increased safety precautions were taken. This included strict safety controls, safe handling of samples and proper disposal of materials after testing. In particular, the hNECs underwent HHH (Triple H: HIV, HCV and HBV) testing to ensure that no contamination occurred during sample collection or experimentation. These tests included sterility testing, viability assessments and contamination testing to ensure the safety and integrity of both the samples and the laboratory environment. After a negative HHH test, the primary cultures can be treated as S1. In addition, the nasal epithelial cells were handled with the utmost care during collection, ensuring that all procedures were performed under sterile conditions to avoid any risk of contaminationFor this purpose, the intensive examination of ethical questions was fundamental and a constant companion of our project. The numerous results from the interviews in the areas of: Ethics, storage and training in the handling of samples have been summarized in a guideline for patient consent for Germany and are intended to provide iGEM teams with the scope, critical examination and observance of iGEM rules, international and national guidelines. 
+ </p>
+ </Collapsible>
+
+ <Collapsible id="CheckDelivery" open={false} title="Check-in for Delivery">
+ <p>
+ Our finished construct is designed to be delivered into the lung via an inhaler using lipid nanoparticles (LNPs). To be more spezific a selective organ-targeting (SORT)- LNPs were developed to deliver mRNA specifically to the lung, with special measures taken to increase biocompatibility and safety. Since the LNP composition is very specific and also differs from other formulas, we submitted the LNP as a checkin:
+ </p>
+ <p>
+ <strong>LNP:</strong> These LNPs are then taken up by epithelial cells through endocytosis, releasing the construct into the cytosol. We carefully evaluated the potential risks, including unintended immune responses and the need for precise dosing to minimize side effects. In addition, we have conducted an in-depth analysis of the dual-use potential of our technology. Dual-use refers to the possibility that scientific advances can be used for both civilian and military purposes. Therefore, we have implemented strict safety protocols and ethical guidelines to ensure that our technology is used exclusively for peaceful and therapeutic applications.
+ </p>
+ </Collapsible>
+ </div>
+ </Section>
+ <Section title="Our Lab" id="Our Lab">
+ <p>
+ As part of our laboratory activities for our <PreCyse/> project, we worked in various laboratories. For general lab work and cloning experiments, you can find some pictures of our laboratories below:
+ </p>
+ <H4 text="Our Cloning Lab"></H4>
+ <p>
+ Our Cloning-laboratory is divided into different work areas to ensure that the experiments run smoothly and efficiently. These include the gel station, the PCR station, the transformation section and the measurement area. Each area is specially equipped for the respective method, and the corresponding experiments were carried out exclusively in the designated stations. In this way, we ensure that our work is carried out under optimal conditions and with the greatest possible precision.
+ <div className="figure-wrapper">
+ <figure>
+ <img src="https://static.igem.wiki/teams/5247/photos/biosafety/kollage/new/img-2041.jpeg" style={{height: "10%", width:"auto"}}/>
+ <figcaption> <b>Figure 1</b> Photo-gallery of laboratory. A: Key lock. B: Key-locked door. C: Alarm plan. D: Emergeny button for electriotion stop. E: Emergency telephone. F: First aid kit, cardiac defibrillaton and emergency exit and fire alarm plan. G: Wash bin with emergency eye wash. H: Emergency shower. I: Lockable cabinets for chemical storage. </figcaption>
+ </figure>
+ </div>
+ <div className="figure-wrapper">
+ <figure>
+ <img src="https://static.igem.wiki/teams/5247/photos/biosafety/kollage/new/img-2037.jpeg" style={{height: "10%", width: "auto"}}/>
+ <figcaption> <b>Figure 2</b> Photo-gallery of S1 laboratory. A: Autoclave. B: Refrigerator with chemicals. C: Weighing room with chemical storage. D: Clean bench work space with vortex, pipettes, heat block and bench top centrifuge. E: pH electrode in fume hood. F: Ice machine. G: Fire distinguisher and S1 waste. H: Fume hood with liquid waste.</figcaption>
+ </figure>
+ </div>
+ </p>
+ <H4 text="Our Cell Culture Lab "></H4>
+ <p>
+ In our cell culture laboratory, we work under sterile conditions to ensure optimal growth conditions for human cell lines. Among other things, we carry out transfections in order to introduce genetic material into cells and investigate their behavior. Strict protocols and state-of-the-art technology ensure the precision and reproducibility of our experiments.
+ </p>
+ <div className="figure-wrapper">
+ <figure>
+ <img src="https://static.igem.wiki/teams/5247/photos/biosafety/kollage/new/img-2040.jpeg" style={{height: "10%", width: "auto"}}/>
+ <figcaption> <b>Figure 3</b> Photo-gallery of laboratory and chemical storage. A: Safety cabinets. B: Incubator. C: Safety cabinet.</figcaption>
+ </figure>
+ </div>
+ <p>
+ In our S2 laboratory, the harvested nasal epithelial cells that serve as primary cultures undergo a comprehensive HHH test to ensure their safety and suitability for further experiments. This test is crucial to ensure that we can subsequently work safely with these cells in the S1 range without the risk of contamination or unwanted release of biological material.
+ </p>
+ <div className="figure-wrapper">
+ <figure>
+ <img src="https://static.igem.wiki/teams/5247/photos/biosafety/kollage/new/img-2042.jpeg" style={{height: "10%"}}/>
+ <figcaption> <b>Figure 4</b>Photo-gallery of S2 laboratory. A: Door of S2 lab with S2 sign. B: Emergency shower and fire distinguisher. C: Clean bench with centrifuge. D: Incubator. E: Safety cabinet. F: Emergeny telephone. G: S2 lab coat with S2 sign. H: Microscope. I: Autoclave. </figcaption>
+ </figure>
+ </div>
+ </Section>
+ </Section>
<Section title="References" id="References">
<ol>{/*<!-- Citation num 1--> */}
<li typeof="schema:ScolarlyArticle" role="doc-biblioentry" property="schema:citation" id="desc-1">
diff --git a/src/data/hptimelinedata.tsx b/src/data/hptimelinedata.tsx
index 74a84e163050675607c767d6512d7fd2ebb6983d..f7a57d00e81a5b1b75e24236f50a959d91e6aa2a 100644
--- a/src/data/hptimelinedata.tsx
+++ b/src/data/hptimelinedata.tsx
@@ -47,7 +47,7 @@ export interface TimelineDatenpunkt {
job?: string;
cardtext: string;
language?: Language;
- quote?: string;
+ quote?: string | Array<string> | Array<React.ReactNode>;
quoteVorname?: string; /* Wenn die quote nicht von der Person ist über die der Text ist */
quoteNachname?: string;
aimofcontact?: string | Array<string> | Array<React.ReactNode>; /* Sobald Zitierungen drin sind oder Links muss es HTML Code sein, ansonsten gehen strings */
@@ -119,6 +119,7 @@ const pics: { [key: string]: string } = {
building: "https://static.igem.wiki/teams/5247/photos/hp/buildingtheteam.webp",
ideas: "https://static.igem.wiki/teams/5247/photos/hp/pitchingideas.webp",
labsupply: "https://static.igem.wiki/teams/5247/photos/hp/labsupply-info.webp",
+ hamburg: "https://static.igem.wiki/teams/5247/photos/hp/gruppenbild-hamburg.webp",
};
@@ -192,7 +193,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
{
vorname: "Ideation in Hamburg",
nachnname: "",
- pictureurl: pics['placeholder'],
+ pictureurl: pics['hamburg'],
job: "Team iGEM",
affiliation: "Bielfeld CeBiTec 2024",
tag: "Milestone",
@@ -220,6 +221,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "maxfirst",
cardtext: "",
language: "de",
+ quoteNachname:"Beckmann, Patient",
+ quoteVorname: "Max",
quote: "A friend of mine with cystic fibrosis recently got a fungal infection that he can't get rid of. His situation really struck me; it showed how quickly a seemingly minor issue can worsen a person's life, especially for someone like us. It’s a strong reminder of how fragile our health is and how fast things can change without warning.",
aimofcontact: [<p>When CF came up as a possible topic, we reached out to a teammate's friend Max in the hopes of getting insights into the needs of CF patients and current treatments to verify the need for further treatment options.
Since he was much more enthusiastic and open for discussion than we dared to hope, we extended our exchanges into the realms of the reality of life for CF patients, possible progressions, organizations and doctors in our area and his personal perspectives and values.
@@ -322,6 +325,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "kristian",
cardtext: "",
language: "de",
+ quoteNachname: " Müller, Reasearch Expert for Gene Therapy ",
+ quoteVorname: "Prof. Dr. Kristiann",
quote: "AAVs have been widely used in gene therapy, but their scalability and immunogenicity pose challenges, particularly when re-dosing is required. In contrast, LNPs offer a promising alternative, with a higher packaging capacity and lower immunogenicity. While AAVs excel in targeted delivery, LNPs could become a more scalable and cost-effective solution for delivering gene-editing tools in the future.",
aimofcontact: [<p>The aim of our interview with Prof. Dr. Kristian Müller was to gain expert insights into the potential of prime editing technology and its application in gene therapy, particularly for treating CF. We sought to understand how innovations in prime editing, combined with optimized delivery systems, could enhance therapeutic outcomes. Prof. Müller’s expertise in molecular biology and gene editing provided valuable perspectives on the opportunities and challenges in developing precise, efficient, and safe treatment strategies for genetic disorders.</p>],
insights: [<p>During the interview, Prof. Dr. Müller highlighted the growing importance of prime editing as a powerful tool for gene therapy. Unlike traditional CRISPR-Cas systems, which often result in double-strand DNA breaks, prime editing allows for precise single-strand cuts, minimizing off-target effects and enabling more specific genetic corrections. This technology opens up new possibilities for treating diseases with known mutations, such as cystic fibrosis.
@@ -360,15 +365,14 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "labsupply",
cardtext: "",
quote: "LabSupply in Münster was a fantastic opportunity to connect with passionate professionals and explore cutting-edge research tools. We're excited about the potential collaborations ahead!",
- quoteNachname: "Sanfilippo",
+ quoteNachname: "Sanfilippo, Teammember",
quoteVorname: "Liliana",
summary: "In April our team visited LabSupply in Münster, where we had the chance to connect with many welcoming and knowledgeable professionals in the lab equipment field. It was an excellent opportunity to explore the latest advancements in laboratory supplies and tools that can enhance our research.",
months: "April",
pictureurl_aim: "https://static.igem.wiki/teams/5247/photos/hp/labsupply.webp",
},
{
- title: "Prof. Dr.",
- vorname: "Wolf-Michael Weber",
+ vorname: "Prof. Dr. Wolf-Michael Weber",
nachnname: "und Dr. Jörg Große-Onnebrink",
job: "Professor ",
affiliation: "Univesity Münster",
@@ -378,7 +382,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "weber",
cardtext: "",
language: "en",
- quote: "x",
+ quote: [<p>This is also an <b>innovative step</b> that you have developed yourselves. That's the part that's really <b>new and significant</b>. A clever and exciting approach with <b>great potential</b>.</p>],
aimofcontact: [<p>The aim of the interview was to gain expert insights on optimizing the delivery of CFTR-mRNA via lung-targeted lipid nanoparticles (LNPs) for cystic fibrosis (CF) treatment.
Specifically, the goal was to explore potential cell targets, challenges in delivery mechanisms, and technical tools for assessing the effectiveness of mRNA therapies like the Ussing chamber system. </p>],
insights: [<p>The experts highlighted the potential of targeting ionocytes, given their key role in CFTR expression, but emphasized the difficulty in accessing them due to their basal position in the respiratory epithelium.
@@ -410,6 +414,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "olariu",
cardtext: "",
language: "de",
+ quoteNachname: "Olariu, Clinical Physician and CF Expert",
+ quoteVorname: "Dr. Cristian-Gabriel",
quote: "For most families, it’s a shock. Cystic fibrosis still has a strong association with being a life-threatening disease, despite the fact that we now have good treatments, and many patients can live healthy lives. The diagnosis puts a huge psychological strain on the family, especially when dealing with very young children.",
aimofcontact: "To gain a deeper insight into the path to diagnosis, we invited pediatrician Dr. Cristian-Gabriel Olariu from the University Department of Pediatrics and Adolescent Medicine to share his experiences with CF patients with us. We interviewed him because of his expertise in the effects of diagnosis on the patient and the family members, but also on daily life. Additionally, we want to close the gap and create a bridge between academic research and clinical applications. Therefore, Dr. Olariu gave us insights about the clinical perspectives on CF patients.",
insights: [<p>We invited Max, our CF patient contact, to join Dr. Olariu in discussing the intersection of academic research, clinical application, and patient needs. Through our connection with <a href="https://www.cfvww.org" >CF Vests Worldwide</a>, an organization dedicated to providing life-saving therapy vests to cystic fibrosis patients globally, we gained insights into the challenges faced by CF patients, particularly in regions like Thailand, where access to advanced treatments and medical devices is limited. The conversation highlighted the critical role of early diagnosis and intervention, as well as the quality-of-life challenges many patients endure due to conventional treatments that may not be effective for everyone. Innovative approaches, such as our SORT LNP (lipid nanoparticle) delivery system, present promising alternatives for CF therapy. This system, which allows for RNA encapsulation and administration via dry spray inhalation, could revolutionize treatment by targeting lung cells more effectively, particularly in resource-limited settings. Dr. Olariu underscored the need for psychological support and coordinated care for CF patients, emphasizing that novel therapies like LNP-based gene treatments have the potential to improve treatment efficacy and accessibility, ultimately reducing the lifelong burden of care for patients and their families. </p>,
@@ -485,6 +491,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "westhoffinv",
cardtext: "",
language: "de",
+ quoteNachname:"Westhoff, Physiotherapist",
+ quoteVorname: "Katrin",
quote: "The more we know, the more opportunities we have.",
aimofcontact: "The objective of the contact was to gain in-depth insights into the treatment and care of children with CF. The therapist's expertise was intended to help develop a better understanding of the challenges and necessary measures in the treatment of this chronic disease. In addition, the aim was to ascertain how the therapy is implemented in everyday life and which specific approaches and methods are particularly effective.",
insights: "The interview yielded valuable insights into the regular implementation of the therapy, the use of aids and the adaptation of exercises to the individual needs of the patients. It was notable that the therapy has improved over the last years, considerably thanks to better medication and adapted exercises, with a concomitant increase in life expectancy for children affected by CF. Of particular interest was the emphasis on the importance of sport and exercise, which should not only be therapeutically effective, but also increase quality of life. ",
@@ -527,7 +535,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
quote: "x",
aimofcontact: "Our primary goal in reaching out to her was to gain insights into optimizing cell culture media, explore potential sponsorship opportunities for our project, and seek expert advice on handling primary cells in the lab.",
insights: "Throughout our multiple discussions, she offered detailed advice on selecting and optimizing cell culture media for our specific needs, while also connecting us with opportunities for sponsorship. Later in the project, she provided technical guidance on the cultivation and handling of primary cells, ensuring that we followed best practices for cell culture.",
- implementation: "Based on her feedback, we refined our approach to cell culture media, enhancing our experimental workflows. Additionally, her advice was critical during the transition to primary cell culture, helping us secure the necessary resources and expertise for our experiments.",
+ implementation: "Based on her feedback, we refined our approach to cell culture media, enhancing our experimental workflows.Her technical support, as well as the support of Julia Watson helped us to establish our experiments in cellculture. Additionally, her advice was critical during the transition to primary cell culture, helping us secure the necessary resources and expertise for our experiments.",
summary: "We have had the privilege of collaborating with a dedicated expert from Stemcell Technologies, who has consistently supported iGEM Bielefeld. She provided valuable guidance on cell culture media, sponsorship opportunities, and later, practical advice for cultivating primary cells. Her contributions have been instrumental in advancing our project.",
months: "several times",
pictureurl_interview: "https://static.igem.wiki/teams/5247/photos/hp/daniela.webp",
@@ -593,6 +601,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "mattijsinv",
cardtext: "",
language: "en",
+ quoteNachname: "Bulcaen, CF Research Expert and Gene Therapy Specialist",
+ quoteVorname: "Mattijs",
quote: "[…] Prime Editing system is more complex than the canonical CRISPR systems, with more variables that can influence success or failure.",
aimofcontact: [<p>Shortly after we decided to use prime editing as the gene editing method for our CF therapy, Mattijs Bulcaen from the Laboratory of Molecular Virology and Gene Therapy at KU Leuven and his colleagues published a paper directly related to our research <TabScrollLink tab="mattijsinv" scrollId="desc-1" num="1" />. In contrast to our approach, Bulcaen et al. 2024 targeted other, less common but drug-refractory CFTR-specific mutations (L227R- and N1303K).  </p>],
insights: [<p>The insights gained from this interview proved extremely valuable in shaping our subsequent mechanistic approaches. At that point in the project we were starting to design the components of our prime editor, but we were lacking a broader overview over the state of the field. Mattijs gave us this insight, mentioning techniques like PE3b systems, dsgRNAs and a talk given by <HPLinktoOtherHPTab tab="liu" text="David Liu" />, the principal
@@ -708,6 +718,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "julia",
cardtext: "",
language: "de",
+ quoteNachname: ", Mother of a CF patient",
+ quoteVorname: "Julia",
quote: "At first, our world fell apart. I still remember the conversation with the doctor. ",
aimofcontact: [<p>We learned from our discussion with <HPLinktoOtherHPTab tab="maxfirst" text="Max" /> that CF has a profound impact on the whole family – not just the patient. In order to gain further insight into this subject, we sought to engage with the next of kin of CF patients.
We were able to make contact with Julia through the self-help group of <a href="https://www.muko.info/ " >Mukviszidose e.V. </a> of which Max is a member. She subsequently reached out to us following Max's request for potential candidates for an interview with a patient group.
@@ -897,7 +909,9 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "radukic",
language:"de",
cardtext: "",
- quote: "x",
+ quote: "After the interview with Dr Marco Radukic, we were able to produce LNPs for the first time.",
+ quoteNachname: "Wiesner, Teammember",
+ quoteVorname: "Lisa",
aimofcontact: [<p>The primary objective was to tackle challenges in LNP transfection related to manufacturing and cell transfection methods. The focus was on improving LNP formulation and application protocols to enhance gene delivery effectiveness,
and on acquiring specialized expertise to optimize these processes. Dr. Radukic from Bielefeld University provided crucial insights for troubleshooting and protocol optimization to enhance LNP efficacy. </p>],
insights: [<p> Dr. Radukic told us that the efficiency of LNPs is significantly affected by lipid-to-nucleic acid ratios and that optimizing ratios like 22:1 versus 10:1 can improve transfection.
@@ -921,7 +935,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "patchclamp",
cardtext: "",
language: "de",
- quoteNachname: "Guckes",
+ quoteNachname: "Guckes, Teammember",
quoteVorname: "Isabell",
quote: "Initially we hadn't considered patch-clamp measurements in our set of downstream applications, but it’s proven to be an exceptionally sensitive method for assessing CFTR conductance.",
aimofcontact: [<p>As part of our project, we aimed to demonstrate the functionality of the CFTR ion channel, after restoring
@@ -1046,7 +1060,9 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "hammer",
cardtext: "",
language: "de",
- quote: "x",
+ quote: "Implementing controls is crucial for the Nikase assay to prevent false positives and ensure the accuracy of your results.",
+ quoteNachname: "Hammer, Expert for Enzyme Engineering ",
+ quoteVorname: "Prof. Dr. Stefan",
aimofcontact: [<p>After we had developed a number of theroetic elaborations, feedback with corresponding expertise in the field of enzyme engineering was of exceptional importance to us. For this reason, we wanted to discuss our approaches with Prof. Dr. Hammer.</p>],
insights: [<p>In our discussion with Prof. Dr. Hammer discussion with Prof. Dr. Hammer about different theoretical approaches we had developed:</p>,
<ul>
@@ -1184,7 +1200,9 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "mattijsvisit",
cardtext: "",
language: "en",
- quote: "x",
+ quote: "It was impressive to meet him in person and realize how deeply knowledgeable he is on the subject; his insights were incredibly helpful.",
+ quoteNachname: "Mundt, Teammember",
+ quoteVorname: "Philip",
aimofcontact: [<p>After our first interview with Mattijs Bulcaen we stayed in contact via email and eventually visited him in Leuven at his laboratory. Here we wanted to gain further information about the CFTR F508del models and editing confirmation.</p>],
insights: [<p>We spoke about approaches for testing CFTR F508del correction in models and methods of confirmation. In this context we talked about HEK293T cell lines[Link] established in his laboratory that stably overexpress CFTR wild-type and F508del. We also discussed how to handle these cell lines. He explained, that the CFTR is fused with a 3HA tag, that in wild-type CFTR would be exposed to the extracellular space and therefore can be used for immunohistochemical staining of the protein, showing correct protein processing and channelling. Also, this allows for a western blot to be made using 3HA antibodies. Functional recovery of CFTR can also be visualized using halide sensitive eYFP or organoid assay, the ladder Mattijs had established an automated readout. Furthermore, we talked about how to handle Sanger sequencing data to analyse edits and discussed the possibility to avoid the weaknesses of Sanger sequencing by using Nanopore sequencing instead. We asked about the applicability of patch clamp analysis in the context of CFTR and Mattijs said that, to his knowledge, it has not been used to test for successful editing in CFTR.
Lastly Mattijs explained how he plans to deliver the prime editing complex to the patient, and we evaluated the advantages and disadvantages of delivery strategies, including our very own LNP approach.</p>],
@@ -1233,7 +1251,9 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "hammerkai",
language: "de",
cardtext: "",
- quote: "x",
+ quote: "It was fascinating to discuss enzyme engineering with a former iGEMer; the conversation sparked new ideas and perspectives.",
+ quoteNachname: "Michalek, Teammember",
+ quoteVorname: "Christian",
aimofcontact: [<p>When we realized that the creation of a nickase from the endonucleases in use was a desired outcome, it became necessary to talk to an expert in the field of enzyme engineering. Our first contact was Kai Schülke, a former iGEMer and PhD student under the guidance of <HPLinktoOtherHPTab tab="hammer" text="Prof. Dr. Hammer"/>, who is the leader of the working group organic chemistry and bioanalytics at Bielefeld University.</p>],
insights: [<p>In the process of our interaction with Kai, we learned about the various methods employed in enzyme engineering. He demonstrated the complexity of this field of research and emphasized the importance of choosing the right approach. As a former iGEMer, Kai, inspired by his past experiences, is highly motivated and determined to develop an outstanding project. He pointed out that we cannot rely on classical methods such as directed evolution, but instead should use a rational approach to select mutation candidates. His insights and enthusiasm have encouraged us to think critically and pursue innovative solutions in our work. </p>],
implementation: [<p>We incorporated Kai's insights into our project by shifting our approach to enzyme engineering. By focusing on a more targeted approach, we were able to refine our enzyme optimization process, ensuring that the modifications we made were based on informed, calculated decisions. This not only streamlined our research but also improved the chances of success by reducing the trial-and-error inherent in traditional methods. </p>],
@@ -1252,7 +1272,9 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
heading: "Adapting expression strategies for Fanzor nickases and exploring the potential of Pichia pastoris for SpuFz1 nickase variants ",
interviewtabid: "nberelsmann",
cardtext: "",
- quote: "X",
+ quote: "Selection for multiple plasmid integrations is the easiest way to optimize protein expression in the yeast strain",
+ quoteNachname:"Berelsmann, Specilist of yeast culturvation",
+ quoteVorname: "Nils",
aimofcontact: [<p>During our interview with <HPLinktoOtherHPTab tab="saito" text="Makoto Saito"/> about fanzor[link fanzor], it became evident that the expression of our fanzor nickases in yeast is very promising. We then refined our expression strategy for the nickases and approached Nils Berelsmann, who is currently working on his PhD thesis with the yeast strain Pichia pastoris (SMD1163). This particular strain could be ideal for expressing the SpuFz1 nickase variants. Our main aim in contacting Nils was to gain insight and advice on yeast expression and he generously shared his expertise with us. Not only did he give us valuable advice, but he also provided us with the yeast strain itself, along with a corresponding expression vector for possible experiments. He also provided us with detailed protocols and the plasmid map of the vector and gave us practical tips on how to optimize the expression process. His support was invaluable in moving our work forward. </p>],
insights: [<p>Pichia pastoris (SMD1163) is a promising option for expressing SpuFz1 nickase variants. Refining expression strategies based on expert insights is crucil for success. Nils provided practical tips on yeast expression, including optimizing growth conditions and fine-tuning induction protocols.</p>],
implementation: [<p>We adapted our expression strategy for Fanzor nickases in yeast by incorporating the Pichia pastoris strain (SMD1163) and the provided expression vector into our experiments. Following Nils' detailed protocols and plasmid map, we optimized key steps, enhancing expression efficiency and protein yield.</p>],
@@ -1270,6 +1292,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
heading: "Discussed the processes involved in the storage, processing, and security of patient samples.",
interviewtabid: "timm",
cardtext: "",
+ quoteVorname:"Dr. Timm",
+ quoteNachname: "Weber, Scientist Project- and Quality Management Biobank",
quote: "A biobank is not just a collection of samples; it's a bridge between patient trust and scientific discovery, ensuring that valuable biological data is safeguarded while contributing to future research.",
aimofcontact: "Contact was established with Timm for the purpose of gaining deeper insights into the functioning of the biobank and of deepening our understanding of the processing of patient samples.",
insights: "We were provided with invaluable insights into the quality and project management of the biobank and storage of patient samples. It was of particular interest to note that Biobank OWL occupies a distinctive position in this context, insofar as a trustee is not a mandatory figure within its system and is therefore not provided for as a standard component. However, Biobank OWL has elected to integrate a trustee in order to enhance the security standards for the safeguarding of patient data. This illustrates the biobank's dedication to ensuring the optimal protection and security of sensitive patient data.",
@@ -1427,6 +1451,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "kolonkofirst",
cardtext: "",
language: "de",
+ quoteVorname:" Dr. Katharina",
+ quoteNachname: "Kolonko, LNP Specialist with focus on chitosan and CF therapies ",
quote: "Wow, you’re already further along than I was! That’s a really good approach, especially since dry powder formulations can help with stability.",
aimofcontact: [<p>Our goal in reaching out to Dr. Katharina Kolonko, who earned her PhD working on chitosan-based nanoparticles for delivering nucleic acids to human respiratory cells in the context of cystic fibrosis, was to seek her advice on the design, stability, and application of nanoparticles. We wanted to learn from her experience with chitosan-capsaicin nanoparticles, especially the challenges she encountered, and apply her insights to improve our own project. Specifically, we aimed to better understand nanoparticle stability, transfection methods, and how to effectively design our experiments.
</p>],
@@ -1490,7 +1516,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
cardtext: "",
quote: "The GXP course was extremely useful as it provided us with important knowledge that supports our entire team in complying with quality standards. This knowledge will help us to organise our processes efficiently and in accordance with regulations in the future.",
quoteVorname:"Kaya",
- quoteNachname:"Lange",
+ quoteNachname:"Lange, Teammember",
type: "meta",
text: [<p>I, Kaya, Team Member of iGEM Bielefeld 2024, recently participated in an intensive one-week GXP (Good Practice) training course, which was pivotal experience for both me and our project. The course covered essential regulatory frameworks, including</p>,
<ul>
@@ -1543,7 +1569,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
cardtext: "",
quote: "The implementation of the hygiene concept is proving more difficult than expected due to the bureaucracy at the university. Nevertheless, the interview gave us a good insight into this labyrinth of regulations and we got started the prozess of implementation.",
quoteVorname: "Vera",
- quoteNachname: "Köhler",
+ quoteNachname: "Köhler, Teammember",
aimofcontact: [<p>We contacted the university because of the urgent need to address the issue of hygiene for students and staff, particularly those with immunocomprised students and staff. There was a need to develop an effective hygiene concept to ensure the health and safety of these people. We developed this concept in collaboration with <HPLinktoOtherHPTab tab="maxfirst" text="Max" />, our CF friend. </p>],
insights: [<p>We learnt that our hygiene concept is very well-developed. But although a well-developed hygiene concept is already existing, strategic development and a step-by-step approach are needed. In particular, the step-by-step implementation was emphasized, like starting with equipping the toilets. Bureaucratic hurdles, such as the need to apply to the rectorate, were identified as a major challenge. In addition, it became clear that there is a great need for sanitary facilities and facilities for the disabled, especially due to the needs of students and staff with health problems. Interaction and networking with other universities was also considered valuable. </p>],
implementation: [<p>The next phase of developing a new hygiene concept is to maintain contact with Mr. Johannfunke in order to continue to advance the hygiene concept in collaboration. The strategic approach entails the incremental implementation of measures, exemplified by the establishment of the inaugural toilet facility within the main building. It is of the utmost importance to ensure the uninterrupted implementation of the hygiene concept. In order to achieve this, it is essential to draw upon the existing plans and measures that have already been implemented in new buildings. We are working on advancing the plans at a higher level and are in regular dialogue with the Central contact point Barrier-free in order to overcome bureaucratic hurdles and actively promote the topic. Furthermore, it is necessary to intensify lobbying work in order to gain greater support for this issue at both the university and political levels. </p>],
@@ -1618,7 +1644,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
months: "several times"
},
{
- title: "Prof.Dr.",
+ title: "Prof. Dr.",
vorname: "Zoya",
nachnname: "Ingatova and Dr. Nikhil Bharti",
job: "Group Leader",
@@ -1628,6 +1654,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
heading: "Shaping the Future of Genetic Therapy: An interview with Prof. Dr. Zoya Ignatova",
interviewtabid: "ignatova",
cardtext: "",
+ quoteNachname: "Ingatova, CF Research Experts ",
+ quoteVorname: "Prof. Dr. Zoya",
quote: "Precision is key to minimizing side effects and ensuring the safety of your therapy.",
aimofcontact: "We conducted the interview with Prof. Dr. Zoya Ignatova and Dr. Nikhil Bharti from the Institute of Biochemistry and Molecular Biology at the University of Hamburg, seeking to deepen our understanding of their research on cystic fibrosis (CF) and explore additional CF mutations, as well as to learn more about cell culture techniques specific to CF research, since they send us the CFBE41o- cell line. Our aim was also to gather more information about their approaches in CF research, particularly their focus on treating genetic mutations like nonsense mutations, which are highly prevalent in CF.",
insights: [<p>We were struck by Ignatova’s story about founding the iGEM team in Hamburg. Her passion for fostering creativity and innovation in science was inspiring. On a technical level, their advice on cell culture was incredibly practical and immediately useful. Dr. Nikhil Bharti explained how they handle CFBE41o- cells and ALI (air-liquid interface) cell culture. This advice directly addressed the challenges we’ve faced in our own lab, giving us a method to improve our cell culture success rates. During our interview with Prof. Dr. Zoya Ignatova and Dr. Nikhil Bharti, their innovative approach to cystic fibrosis (CF) therapy, particularly "read-through" and tRNA-based therapies, stood out. "Read-through" therapies aim to bypass premature stop codons that prevent full protein production, offering a way to restore the function of critical proteins like CFTR in CF. This approach has the potential to treat a broad range of genetic diseases caused by similar mutations. The tRNA-based therapy is even more precise, targeting mRNA to correct faulty codons without altering the DNA, making it safer for long-term use. This flexibility, along with the ability to apply these therapies beyond CF, broadened our understanding of how such strategies can revolutionize treatments for genetic disorders.Ignatova highlighted, that using cell models closely resembling properties of the cell types targeted by our therapy is important for ensuring applicability of our approach to patient cells and its safety. A key focus throughout the discussion was safety. Prof. Ignatova emphasized the importance of ensuring that the therapies are highly specific, targeting only the defective codons while avoiding natural stop codons to prevent uncontrolled protein production. Moreover, their careful consideration of delivery systems further demonstrated their commitment to minimizing risks like toxicity in unintended organs. Their meticulous approach to safety has influenced how we think about developing and applying these therapies, making it clear that ensuring patient safety is as critical as achieving therapeutic success.</p>],
@@ -1768,7 +1796,9 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
interviewtabid: "saito",
cardtext: "",
language:"en",
- quote: "X",
+ quoteNachname: "Saito, Leading research expert of FANZOR ",
+ quoteVorname: "Dr. Makoto",
+ quote: "This project is actually a really hot, very important project at the absolute cutting edge of science. Honestly, I see researchers around the world working on developing smaller CRISPR-Cas-like prime editors. It shows that you guys are working on a really important problem.",
aimofcontact: "The aim of the interview was to gain deeper insights into the topic of protein engineering, especially with regard to Fanzor (SpuFz) and to get feedback on our existing approaches for possible nickases, as well as for the planned nickase assay. ",
insights: "The interview provided the iGEM team with valuable advice regarding their Prime Editing project and especially on their planned nickase assay. Dr Saito gave detailed feedback on technical challenges, especially with protein expression in E. coli, and suggested switching to yeast for better results. He also encouraged the team to plan carefully, given the project's complexity, and offered guidance on future experiments.  ",
implementation: "We have adapted our planned nickase assay according to Dr Saito's advice and changed it accordingly to expression of the RNP complex using yeast.",
@@ -1814,6 +1844,8 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
heading: "Deep Dive into Good Practise, GxP ",
interviewtabid: "gxpexpert",
cardtext: "",
+ quoteNachname: "Berger, GxP Expert",
+ quoteVorname: "Dr. rer. nat. Marcus",
quote: "Minimum requirement: Acute toxicity study on animals - important for calculating the starting dose for humans. In addition, pharmacological studies, genotoxic studies and immunotoxic studies. In your case also chronic toxicity studies.",
aimofcontact: [<p>The aim of our contact with GxP expert <a href="https://gxpexpert.de/">Dr. Marcus Berger</a> was to gather insights and ask questions about how to proceed with the Precyse project, clarifying the next steps required for its continued development, particularly concerning regulatory strategy, quality management, and clinical development. GxP, which stands for ‘Good Practice’ guidelines, encompasses various regulations and standards intended to ensure quality and compliance in the development and manufacturing of pharmaceutical and biopharmaceutical products. </p>],
insights: [<p>From the discussion with Dr. Berger expert, we gained several key insights into how to proceed with the Precyse project. First, it became clear that obtaining a Scientific Advice from the Paul-Ehrlich-Institut <a href=" https://www.pei.de/DE/home/home-node.html" >(PEI)</a>
@@ -1905,6 +1937,24 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
summary: "In summary, the insights from Benjamin’s expertise were crucial in shaping our understanding of how to integrate Chitosan into our LNP formulations. Chitosan’s protective abilities for RNA, along with its heat stability, make it a valuable component in our project. However, its hydrophilic and cationic nature presents challenges for direct integration into LNP lipid shells, so we are focusing on its use as an encapsulation for the RNA. Benjamin’s advice on concentrations, molecular weight, and complex formation gave us a clear path forward, which will be validated through experimental testing. ",
months: "september"
},
+ {
+ vorname: "Grand Jambooree in Paris",
+ nachnname: "",
+ pictureurl: pics['placeholder'],
+ job: "Team iGEM",
+ affiliation: "Bielfeld CeBiTec 2024",
+ tag: "Milestone",
+ heading: "Brainstorming and selection of ideas and concepts",
+ interviewtabid: "jamboree",
+ cardtext: "",
+ quote: "",
+ aimofcontact: "",
+ insights: "",
+ implementation: "",
+ type: "meta",
+ summary: "",
+ months: "october"
+ },
{
vorname: "Carry It Forward",
nachnname: "",