diff --git a/src/contents/Human Practices/Further Engagement/Collaborations.tsx b/src/contents/Human Practices/Further Engagement/Collaborations.tsx
index e83196abb5f50ae6fdbee3cc94511809e20bf94a..d3e9c1ea39ecfb356364d4f916cf8ba2b24a0d8f 100644
--- a/src/contents/Human Practices/Further Engagement/Collaborations.tsx
+++ b/src/contents/Human Practices/Further Engagement/Collaborations.tsx
@@ -46,9 +46,9 @@ export function HPCollabs(){
<H5 text="Biosafety and Security"/>
<p>Early in our project, we faced challenges working with human biomaterials, particularly cultivating primary human nasal epithelial cells from both CF patients and controls. To address these, we made three key contributions:</p>
<ol>
- <li>A guideline for handling biomaterials in compliance with BSL2 standards.[Link guideline]</li>
- <li>A clinical trial-style questionnaire to assess donor medical history.[Link Link questionaire]</li>
- <li>A hygiene protocol to improve safety and cleanliness in research facilities.[Link hygiene protocoll]</li>
+ <li>A guideline for handling biomaterials in compliance with BSL2 standards.{/* [Link guideline] */}</li>
+ <li>A clinical trial-style questionnaire to assess donor medical history.{/* [Link Link questionaire] */}</li>
+ <li>A hygiene protocol to improve safety and cleanliness in research facilities.{/* [Link hygiene protocoll] */}</li>
</ol>
<p>
@@ -57,12 +57,12 @@ export function HPCollabs(){
</p>
<H5 text="PreCL Reporter System "/>
<p>To test Prime Editing systems targeting the CF-specific delF508 mutation, we developed the PreCL reporter system [Link engineering of PreCL], which offers high sensitivity, minimal noise, and precise fluorescence detection. This versatile tool, adaptable for CRISPR and base editing, enhances the precision of genetic research, particularly in CF studies.
- We optimized pegRNA design[linkpegrna] by incorporating the TevoPreQ1 RNA motif, improving stability and Prime Editing efficiency. Our innovations, including silent edits and fine-tuned sequences, boost editing accuracy, providing a robust tool for genetic research. </p>
+ We optimized pegRNA design{/* [linkpegrna] */} by incorporating the TevoPreQ1 RNA motif, improving stability and Prime Editing efficiency. Our innovations, including silent edits and fine-tuned sequences, boost editing accuracy, providing a robust tool for genetic research. </p>
<H5 text="Prime Editing Technology PrimeGuide & Lipid Nanoparticle System AirBuddy "/>
- <p>Our PrimeGuide[link] system introduces a novel eukaryotic RNA-binding DNA-nickase, a smaller alternative to Cas9. Enhanced with a more efficient Reverse Transcriptase and optimized RNA-binding proteins, this advancement improves Prime Editing accuracy and safety for genetic mutation correction.
- We developed AirBuddy[link], a lung-specific RNA/DNA delivery system optimized for gene therapies targeting lung diseases. With low cytotoxicity, efficient cellular uptake, and cost-effective storage, AirBuddy revolutionizes lung disease treatments by providing a safer and more effective delivery method. </p>
+ <p>Our PrimeGuide{/* [link] */}system introduces a novel eukaryotic RNA-binding DNA-nickase, a smaller alternative to Cas9. Enhanced with a more efficient Reverse Transcriptase and optimized RNA-binding proteins, this advancement improves Prime Editing accuracy and safety for genetic mutation correction.
+ We developed AirBuddy{/* [link] */}, a lung-specific RNA/DNA delivery system optimized for gene therapies targeting lung diseases. With low cytotoxicity, efficient cellular uptake, and cost-effective storage, AirBuddy revolutionizes lung disease treatments by providing a safer and more effective delivery method. </p>
<H5 text="Wiki Development"/>
- <p>To support future iGEM teams, we developed troubleshooting guides for HTML and CSS[link], making wiki development more accessible and easier to manage.
+ <p>To support future iGEM teams, we developed troubleshooting guides for HTML and CSS{/* [link] */}, making wiki development more accessible and easier to manage.
Through these contributions, we provide valuable tools and frameworks to advance synthetic biology, ensuring safer, more efficient research and therapeutic development for the iGEM community. </p>
<H5 text="Global Impact and Inclusivity "/>
<p>Recognizing the disparities in CF care across different regions, particularly in underrepresented areas like Asia, we adjusted our approach to create a more inclusive therapy. With feedback from stakeholders like <a onClick={() => goToPagesAndOpenTab('joshua', '')}>Joshua Bauder</a> from CF Vest International and <a onClick={() => goToPagesAndOpenTab('sriram', '/human-practices')}>Dr. Sriram Vaidyanathan</a>, we ensured our therapy addressed a wider range of CF mutations. This global focus led to bilingual surveys and expanded outreach efforts to raise awareness about CF and gene therapy. </p>
diff --git a/src/contents/parts.tsx b/src/contents/parts.tsx
index a842b0941b4d16976fab2267afc3d62b297634ea..e935c1f9895221165555d258521486e3e1b9c5e2 100644
--- a/src/contents/parts.tsx
+++ b/src/contents/parts.tsx
@@ -58,9 +58,9 @@ export function Parts() {
<p>In the next series of experiments, we would like to investigate various mutation candidates, in particular the possible SpuFz1 nickases (BBa_K5247101- BBa_K5247104) and various PlmCasx nickase variants (BBa_K5247105- BBa_K5247107), in more detail using the PE6c system. </p>
{/* Bild Nikase */}
<H4 text="nSpuFz1 "/>
- <p>The nSpuFz1 variants are expressed in yeast strain Pichia pastoris (SMD1163), which we obtained from Nils Berelsmann[link zu HP-Timeline]. In advance, the corresponding genes were cloned into a suitable expression vector, pPIC9K, via Gibson Assembly to ensure efficient expression of the nickases. The cloning as well as the subsequent expression and purification of the nickases were carried out according to a detailed protocol[2] under the expert guidance of Hakan Soytürk[link zu HP timeline]. </p>
+ <p>The nSpuFz1 variants are expressed in yeast strain Pichia pastoris (SMD1163), which we obtained from Nils Berelsmann{/* [link zu HP-Timeline]. */} In advance, the corresponding genes were cloned into a suitable expression vector, pPIC9K, via Gibson Assembly to ensure efficient expression of the nickases. The cloning as well as the subsequent expression and purification of the nickases were carried out according to a detailed protocol[2] under the expert guidance of Hakan Soytürk[link zu HP timeline]. </p>
<H4 text="nPlmCasX "/>
- <p>The nPlmCasX variants are expressed in E. coli strain BL21D3, which we obtained from AG Müller[link zu HP-Timeline]. In advance, the corresponding genes were cloned into a suitable expression vector, pZMB1029, via Gibson Assembly to ensure efficient expression of the nickases. The cloning as well as the subsequent expression and purification of the nickases were carried out according to a detailed protocol[2]. </p>
+ <p>The nPlmCasX variants are expressed in E. coli strain BL21D3, which we obtained from AG Müller{/* [link zu HP-Timeline] */}. In advance, the corresponding genes were cloned into a suitable expression vector, pZMB1029, via Gibson Assembly to ensure efficient expression of the nickases. The cloning as well as the subsequent expression and purification of the nickases were carried out according to a detailed protocol[2]. </p>
{/* Bild beide */}
<p>After successful purification, the isolated nickases are comprehensively analyzed according to verify their activity and efficiency. These analyses will serve to evaluate the functionality and suitability of the nickases for specific applications in prime editing. Subsequently, detailed characterization experiments are planned to determine the properties of the nickases, if functional, including their specificity, editing activity and potential for use in precise gene editing procedures. </p>
<p>Validation of the nickases will be performed in different cell lines to confirm their efficiency and reliability in a cellular context. These validation steps are crucial to further investigate the potential of Prime Guide for therapeutic applications. </p>
diff --git a/src/contents/results.tsx b/src/contents/results.tsx
index ea76fcfe1951db6040c7cd1ac657657376a2378b..2b59477831f458d0e18f15425573803533483438 100644
--- a/src/contents/results.tsx
+++ b/src/contents/results.tsx
@@ -331,7 +331,7 @@ export function Results() {
</figure>
</div>
</div>
- <p>Overall, while the Cryo-EM data confirm the presence and general morphology of LNPs that also fall within the diameter range specified by Wang et al. for SORT LNPs at smaller than 200 nm [link]. The variability in size and the presence of aggregates highlight potential areas for optimization, such as refining sample concentration and preparation methods to achieve more consistent particle formation.</p>
+ <p>Overall, while the Cryo-EM data confirm the presence and general morphology of LNPs that also fall within the diameter range specified by Wang et al. for SORT LNPs at smaller than 200 nm {/* [link] */}. The variability in size and the presence of aggregates highlight potential areas for optimization, such as refining sample concentration and preparation methods to achieve more consistent particle formation.</p>
<H5 text="DLS"/>
<p>We used Dynamic Light Scattering (DLS) to assess the size distribution of our SORT LNPs by measuring the fluctuations in scattered light due to particle motion. The hydrodynamic diameter was calculated using the Stokes-Einstein equation, considering the diffusion coefficient, temperature, and viscosity of the medium.</p>
<div className="row align-items-center">
diff --git a/src/contents/safety.tsx b/src/contents/safety.tsx
index 15c47f010a478cd8edf68ebdb30bc74502cec049..c35a2c68b192648433fd44ae8857934bce3faffa 100644
--- a/src/contents/safety.tsx
+++ b/src/contents/safety.tsx
@@ -30,7 +30,7 @@ export const Safety: React.FC = () =>{
</p>
<H4 text="PAM disrupt" ></H4>
<p>
- A key safety mechanism incorporated in our design of the Prime Editing complex is the disruption of the PAM sequence [Link PAM text]. For the nickase enzyme to function properly, it must bind directly to the DNA strand, a process that is facilitated by the presence of a specific sequence called the PAM (Protospacer Adjacent Motif). This critical interaction occurs through the recognition of the PAM sequence by the nickase itself. To achieve PAM disruption, the pegRNA (prime editing guide RNA) [Link pegRNA] is specifically designed in a way so that the PAM sequence is situated within the reverse transcription template (RTT) of the pegRNA. By introducing a silent mutation within the RT template into the PAM sequence. Therefore the PAM sequence is effectively eliminated after the gene editing process is successfully completed <SupScrollLink label="1"/>. As a result of that, the PAM sequence is no longer present on the DNA strand, preventing the nickase from binding again at the same location. This reduction in repeated or undesired binding of the nickase enhances the safety of our prime editing complex, minimizing the risk of unintended edits or off-target effects in subsequent steps. Ultimately, this feature contributes very much to the overall safety and reliability of the prime editing process.
+ A key safety mechanism incorporated in our design of the Prime Editing complex is the disruption of the PAM sequence{/* [Link PAM text] */}. For the nickase enzyme to function properly, it must bind directly to the DNA strand, a process that is facilitated by the presence of a specific sequence called the PAM (Protospacer Adjacent Motif). This critical interaction occurs through the recognition of the PAM sequence by the nickase itself. To achieve PAM disruption, the pegRNA (prime editing guide RNA) [Link pegRNA] is specifically designed in a way so that the PAM sequence is situated within the reverse transcription template (RTT) of the pegRNA. By introducing a silent mutation within the RT template into the PAM sequence. Therefore the PAM sequence is effectively eliminated after the gene editing process is successfully completed <SupScrollLink label="1"/>. As a result of that, the PAM sequence is no longer present on the DNA strand, preventing the nickase from binding again at the same location. This reduction in repeated or undesired binding of the nickase enhances the safety of our prime editing complex, minimizing the risk of unintended edits or off-target effects in subsequent steps. Ultimately, this feature contributes very much to the overall safety and reliability of the prime editing process.
</p>
<H4 text="pegRNA design - Spacer"></H4>
<p>
@@ -64,11 +64,11 @@ export const Safety: React.FC = () =>{
<Subesction title="Safety aspects of our Airbuddy" id="Biosafety2">
<H4 text="SORT LNP and Cytotoxicity"></H4>
<p>
- We have carefully considered the biosafety aspects of our delivery system, starting with the decision between Adeno-associated viruses (AAV) or LNPs as delivery systems. Our comparison revealed that the biocompatibility and safety of LNPs are paramount for our approach. That is why we chose selective organ-targeting (SORT) lipid nanoparticles (LNPs) [Link LNP text] <SupScrollLink label="7"/> in the context of targeted pulmonary mRNA delivery. One of our primary concerns with the LNP was the potential cytotoxicity of polyethylene glycol (PEG), a common stabilizing agent in LNP formulations. Aware of the immune responses PEG can trigger, potentially leading to cytotoxicity <SupScrollLink label="8"/>, we aimed at optimizing its concentration in our SORT LNPs to minimize such reactions while maintaining therapeutic efficacy. By the use of low molecular weight PEG, we addressed this problem. To test weather our approach succeeded, we conducted MTT and proliferation assays to ensure that our LNP posed no cytotoxicity risks.
+ We have carefully considered the biosafety aspects of our delivery system, starting with the decision between Adeno-associated viruses (AAV) or LNPs as delivery systems. Our comparison revealed that the biocompatibility and safety of LNPs are paramount for our approach. That is why we chose selective organ-targeting (SORT) lipid nanoparticles (LNPs){/* [Link LNP text] */}<SupScrollLink label="7"/> in the context of targeted pulmonary mRNA delivery. One of our primary concerns with the LNP was the potential cytotoxicity of polyethylene glycol (PEG), a common stabilizing agent in LNP formulations. Aware of the immune responses PEG can trigger, potentially leading to cytotoxicity <SupScrollLink label="8"/>, we aimed at optimizing its concentration in our SORT LNPs to minimize such reactions while maintaining therapeutic efficacy. By the use of low molecular weight PEG, we addressed this problem. To test weather our approach succeeded, we conducted MTT and proliferation assays to ensure that our LNP posed no cytotoxicity risks.
</p>
<H4 text="Precision of our SORT LNP"></H4>
<p>
- To further improve safety, we focused on reducing off-target effects. By incorporating specific SORT molecules, such as permanently cationic lipids like DOTAP [Link LNP design], we ensured that the nanoparticles are systematically directed to the lungs. This precise targeting is particularly beneficial for respiratory diseases, as it enhances therapeutic effectiveness while limiting the impact on non-target organs. Our outlook of antibody conjugation as surface modification of our LNP for cell type-specific delivery, more exactly club cells <SupScrollLink label="9"/> and ionocytes <SupScrollLink label="10"/> as CFTR-expressing lung epithelial cells, would round off this aspect.
+ To further improve safety, we focused on reducing off-target effects. By incorporating specific SORT molecules, such as permanently cationic lipids like <a onClick={() => goToPageWithTabAndScroll({tabId:'tab-delivery', path: '/engineering', scrollToId: "delivery-header"})}>DOTAP</a> , we ensured that the nanoparticles are systematically directed to the lungs. This precise targeting is particularly beneficial for respiratory diseases, as it enhances therapeutic effectiveness while limiting the impact on non-target organs. Our outlook of antibody conjugation as surface modification of our LNP for cell type-specific delivery, more exactly club cells <SupScrollLink label="9"/> and ionocytes <SupScrollLink label="10"/> as CFTR-expressing lung epithelial cells, would round off this aspect.
</p>
<p>
In summary, our design strategy emphasizes both safety and efficacy. The careful optimization of components like PEG 2000 and the use of targeted delivery molecules allow SORT LNPs to deliver therapeutic agents directly to the lungs, reducing systemic exposure and minimizing side effects. This targeted approach ensures more effective treatments, especially for conditions requiring localized intervention.
@@ -251,7 +251,7 @@ export const Safety: React.FC = () =>{
</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.
+ 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>
diff --git a/src/data/hptimelinedata.tsx b/src/data/hptimelinedata.tsx
index c11aad510ce595d470af99f72576c313b97d623a..328d0441551e5a146b092a734a8dfa9e9f7427ec 100644
--- a/src/data/hptimelinedata.tsx
+++ b/src/data/hptimelinedata.tsx
@@ -252,7 +252,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
“How positively and calmly Max deals with his illness but has also pointed out that he is lucky, and that other people are much worse off - how much you have to pay attention to little things that you wouldn't have expected as a healthy person.†</p>
</>],
implementation: [<> <p>This most important aspect of this meeting was less an insight, but the fact Max helped us to put a face to an abstract idea. Many of our ideas to treat CF were interesting and adventurous but meeting him put a lot into perspective. </p>
- <p>Max played a significant role in shaping our project from the outset, particularly in guiding our focus on Integrated Human Practices [Link Best HP] and Safety & Security [Link Best Biosafety] as special prizes. Through our discussions, we recognized the importance of these aspects in developing the best possible cure. This collaborative effort led us to pivot our target from the pancreas to the lung and move away from a diagnostic approach. His contributions not only provided valuable insights but also fostered a strong personal investment in our project.</p></>],
+ <p>Max played a significant role in shaping our project from the outset, particularly in guiding our focus on Integrated Human Practices {/* [Link Best HP] */}and Safety & Security {/* [Link Best Biosafety] */}as special prizes. Through our discussions, we recognized the importance of these aspects in developing the best possible cure. This collaborative effort led us to pivot our target from the pancreas to the lung and move away from a diagnostic approach. His contributions not only provided valuable insights but also fostered a strong personal investment in our project.</p></>],
pictureurl_implementation: "",
interview: <><QaBox q="How and when were you first diagnosed? " a="When I was about one year old. My mother did not do any screenings or prenatal testing. I was in pain but as an infant you cannot say that, so I screamed a lot. Many doctors shrug that off in small children but after some time a sweat test was done at the children's clinic." />
<QaBox q="What do you think about diagnosing via sweat tests?" a="I am a clear opponent of diagnosing via sweat tests, especially if it is used to rule out CF and people have atypical CF, because of which they do not get diagnosed because of that." />
@@ -335,7 +335,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
Prof. Dr. Kristian Müller emphasized the critical role of delivery systems in the success of gene therapies, particularly in the context of CF treatment. Two primary delivery mechanisms were discussed: AAVs (Adeno-associated viruses) and LNPs (Lipid nanoparticles), each with distinct advantages and limitations.
AAVs are a well-established vehicle in gene therapy, having been used successfully in various approved treatments. They are highly efficient at delivering genetic material to target cells, especially in well-characterized diseases like CF. One of their key strengths is their ability to precisely target specific tissues, making them particularly valuable for lung delivery in cystic fibrosis. However, AAVs come with notable challenges, primarily their limited packaging capacity (approximately 4.5 kilobases), which constrains the size of the genetic payload they can carry. Additionally, AAVs can elicit immune responses, particularly when multiple doses are required, posing a barrier to their long-term use.
On the other hand, LNPs offer a scalable and re-dosable alternative. LNPs have the advantage of a larger packaging capacity, allowing them to carry more complex genetic instructions or larger gene-editing tools, such as prime editors. They are also easier and cheaper to produce on a large scale, making them an attractive option for widespread clinical applications. A significant benefit of LNPs is their lower immunogenicity, which reduces the risk of adverse immune reactions upon repeated dosing. However, LNPs currently face challenges in specific targeting compared to AAVs. AAVs have a higher precision in targeting specific tissues, while LNPs still need optimization for targeted delivery to areas like the lungs.</p>],
- implementation: [<p>Prof. Müller’s insights directly inform the implementation of our iGEM project, where we aim to design novel prime editors that are small enough to be delivered efficiently, while also exploring LNPs[LINK Cycle Delivery] as a scalable and re-dosable alternative to AAVs. By tailoring our approach to address the specific challenges of CF, such as mucus penetration and lung cell targeting, we can enhance the precision and efficacy of gene therapy. These innovations have the potential to set new standards in the field and contribute to broader research on genetic disease treatment.</p>],
+ implementation: [<p>Prof. Müller’s insights directly inform the implementation of our iGEM project, where we aim to design novel prime editors that are small enough to be delivered efficiently, while also exploring LNPs{/* [LINK Cycle Delivery] */} as a scalable and re-dosable alternative to AAVs. By tailoring our approach to address the specific challenges of CF, such as mucus penetration and lung cell targeting, we can enhance the precision and efficacy of gene therapy. These innovations have the potential to set new standards in the field and contribute to broader research on genetic disease treatment.</p>],
summary: "In our interview with Prof. Dr. Kristian Müller, we explored the revolutionary potential of prime editing as a next-generation gene editing technology. Prof. Müller highlighted the advantages of prime editing over traditional CRISPR-Cas systems, particularly its ability to make precise genetic modifications without double-strand breaks, thus reducing off-target effects. He emphasized the importance of optimizing delivery systems, such as AAV and LNPs, and discussed the ethical considerations and biosafety measures crucial for advancing gene therapy. The interview underscored the significance of cystic fibrosis as a model disease, given its prevalence and the potential for impactful treatments through targeted genetic corrections.",
months: "April",
interview:<><iframe title="Bielefeld-CeBiTec: Interview Müller AAV vs LNP (2024) [English]" width="560" height="315" src="https://video.igem.org/videos/embed/0613b6b8-7755-4373-9d86-9910fe30781f" frameBorder="0" allowFullScreen={true} sandbox="allow-same-origin allow-scripts allow-popups allow-forms"></iframe><p>This interview was recorded on video at a later date.</p></>,
@@ -535,7 +535,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
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. ",
implementation: "The following statement by Katrin Westhoff had a particular impact on our project: ‘The more we know, the more options we have’. We learnt from the interview that the current medication is already helping many patients very well, but that there is still great potential for improvement. Successful gene therapy would significantly improve the quality of life of CF patients. We implemented the findings from this interview in our participation in MukoMove - we also actively took part in cystic fibrosis awareness month and learnt even more about the importance of physiotherapy.",
pictureurl_interview: "https://static.igem.wiki/teams/5247/photos/hp/katrin-westhoff-zoom.webp",
- summary: [<p>The objective of our discussion with a therapist was to gain a comprehensive understanding of the treatment and care of children with cystic fibrosis. The interview provided invaluable insights into the therapy's implementation, highlighting the significant advancements in medication and tailored exercises that have led to improved patient outcomes and increased life expectancy. A key takeaway was the emphasis on the role of sports and exercise, not just for therapeutic efficacy but also for enhancing overall quality of life. It let to our participation in the CF awarness month and the outreach project mukomove [Link mukomove]</p>],
+ summary: [<p>The objective of our discussion with a therapist was to gain a comprehensive understanding of the treatment and care of children with cystic fibrosis. The interview provided invaluable insights into the therapy's implementation, highlighting the significant advancements in medication and tailored exercises that have led to improved patient outcomes and increased life expectancy. A key takeaway was the emphasis on the role of sports and exercise, not just for therapeutic efficacy but also for enhancing overall quality of life. It let to our participation in the CF awarness month and the outreach project mukomove {/* [Link mukomove] */}</p>],
months: "May",
interview:<>
<QaBox q="From what age do the patients come to you? How long do they stay? How many patients do you treat?" a="The patients come to us at a very early age. A definite diagnosis is made after 6 weeks at the latest. Once diagnosed, the whole family is genetically tested, and children are sent for physiotherapy, often starting in the hospital. Currently, we have 8 children with CF in our practice, which is relatively small compared to other diseases. We have slightly more CF patients because we specialize in it."/>
@@ -627,7 +627,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
quoteNachname: "Susat, Teammember",
quoteVorname: "Kathleen",
quote: "It was amazing to see how movement can bring people together. I had a great time.",
- summary: [<p>MUKOmove[LINK unten mukomove] is a sports initiative by Mukoviszidose e.V., the German Cystic Fibrosis Association, to raise awareness and funds for cystic fibrosis. Team iGEM Bielefeld participated from May 8th to May 12th, promoting the event at their university and city, encouraging others to join and collect sport hours. They organized a team event at their university, involving sports games to promote community engagement. Their efforts helped surpass their goal of 240 hours, with the team achieving 358 sport hours, while the entire event gathered over 36,000 sport hours. The initiative successfully raised awareness about cystic fibrosis and promoted physical activity as a means of community building.</p>],
+ summary: [<p>MUKOmove{/* [LINK unten mukomove] */} is a sports initiative by Mukoviszidose e.V., the German Cystic Fibrosis Association, to raise awareness and funds for cystic fibrosis. Team iGEM Bielefeld participated from May 8th to May 12th, promoting the event at their university and city, encouraging others to join and collect sport hours. They organized a team event at their university, involving sports games to promote community engagement. Their efforts helped surpass their goal of 240 hours, with the team achieving 358 sport hours, while the entire event gathered over 36,000 sport hours. The initiative successfully raised awareness about cystic fibrosis and promoted physical activity as a means of community building.</p>],
months: "May"
},
{
@@ -647,7 +647,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
quote: "The interview proved to be invaluable in gaining an initial understanding of the principles of pegRNA design and optimisation, particularly in the context of silent edits.",
aimofcontact: "The aim of the contact was to engage in a discussion about prime editing and pegRNAs, as the Jan-Phillip Gerhards had used these technologies in his internship at the Boston Childrens Hospital. We sought to exchange ideas, gather insights, and explore potential improvements or strategies for our project, leveraging his experience with prime editing tools. His practical knowledge in this field was very valuable for refining our approach and ensuring we were aligned with the latest advancements and methodologies in prime editing. ",
insights: "During our discussion we gained valuable insights that had a significant impact on our project. One of the most important findings was the effectiveness of silent edits, which will enable us to make our PrimeGuide safer. Silent edits change the sequence of bases in the DNA in such a way that the resulting protein remains unchanged, because the genetic code is redundant. This means that different codons can code for the same amino acid. By making silent edits in addition to correcting the CFTR gene, we can prevent the pegRNA from rebinding. We have also learned that the length of the primer binding site (PBS) plays a crucial role in determining optimal results and that it is recommended to keep the PBS temperature close to 37°C. Specifically, PBS lengths of 17nt (38.3°C) and 16nt (36.4°C) were found to be ideal options. For our planned set of 12 samples, it was recommended to use three different PBS lengths (differing by +/- 1nt from that close to 37°C) in combination with four reverse transcriptase template (RTTs) to achieve the best result. Another important finding was the use of non-annotated regions with overhangs for cloning, which could give better results in our experiments. However, we also encountered concerns that circRNA, a covalently closed circular RNA molecule, might be sterically hindered by Cas9, which we need to investigate further. When discussing cloning overhangs, we learned that a base-pair length close to 60°C is optimal. However, the use of a 15nt PBS was not recommended as it has a lower temperature range which could affect performance. Although we still need to confirm the oligonucleotide delivery time, these findings will help us to refine our cloning strategy, optimize PBS selection and improve our overall approach to primer editing, especially in terms of the pegRNA design.",
- implementation: "We incorporated the lessons learned from our discussions on prime editing and silent editing directly into our project by refining our approach to gene editing. Based on feedback about the optimal length of primer binding sequences (PBS) and RTTs, we adjusted the design of our pegRNAs to ensure greater precision and efficiency in our experiments. In particular, we learned that using PBS lengths close to 37°C melting temperatures (e.g. 16-17 nucleotides) increased stability, which led us to fine-tune these sequences for improved editing results. The concept of silent editing became an integral part of our safety strategy[Link Biosafety], allowing us to make changes to the DNA more precise. We also revised our cloning strategies by considering the appropriate overhang length, targeting a base pair length near the melting temperature of 60°C to improve cloning efficiency. We also reassessed the practicality of ordering shorter PBS sequences, concluding that lengths shorter than 15 nt were less advantageous due to reduced efficiency. By integrating these findings, we optimised our experimental workflow and made informed decisions about the tools and methods for our prime editing experiments. ",
+ implementation: "We incorporated the lessons learned from our discussions on prime editing and silent editing directly into our project by refining our approach to gene editing. Based on feedback about the optimal length of primer binding sequences (PBS) and RTTs, we adjusted the design of our pegRNAs to ensure greater precision and efficiency in our experiments. In particular, we learned that using PBS lengths close to 37°C melting temperatures (e.g. 16-17 nucleotides) increased stability, which led us to fine-tune these sequences for improved editing results. The concept of silent editing became an integral part of our safety strategy, allowing us to make changes to the DNA more precise. We also revised our cloning strategies by considering the appropriate overhang length, targeting a base pair length near the melting temperature of 60°C to improve cloning efficiency. We also reassessed the practicality of ordering shorter PBS sequences, concluding that lengths shorter than 15 nt were less advantageous due to reduced efficiency. By integrating these findings, we optimised our experimental workflow and made informed decisions about the tools and methods for our prime editing experiments. ",
summary: "We engaged in a valuable discussion with Jan-Phillip Gerhards regarding prime editing and pegRNAs, leveraging his internship experience with these technologies. Key insights included the effectiveness of silent edits, which can enhance the safety of our PrimeGuide by modifying DNA sequences without altering the resultant protein, thereby preventing pegRNA rebinding. We also learned the importance of optimizing the primer binding site (PBS) length to achieve ideal temperatures close to 37°C, recommending lengths of 16-17 nucleotides. Additionally, we discovered the potential benefits of using non-annotated regions with overhangs for cloning, while also recognizing concerns about circRNA steric hindrance by Cas9. These insights directly informed our project, allowing us to refine our pegRNA design and cloning strategies, ultimately enhancing the precision and efficiency of our gene editing approach.",
months: "May"
},
@@ -781,7 +781,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
quoteNachname: "Sahami Moghaddam, Teammember",
quoteVorname: "Asal",
quote: "I had a great time educating the kids about gene therapy and CF in a playful way, and there were some very interesting conversations with the parents.",
- summary: [<p>"Der Teuto ruft!"[LInk zu Teuto unten] is a community event in Bielefeld where iGEM Bielefeld participated to raise awareness about cystic fibrosis (CF) and gene therapy. They engaged children through interactive experiments, such as creating lung models and simulating mucus to help them understand the challenges faced by CF patients. Adults were informed about their gene therapy project and had meaningful discussions about the implications of CF treatment. Collaborating with other institutions like the life science student initiative btS, the team expanded their outreach. Despite changeable weather, the event was a success in educating the public and improving science communication.</p>],
+ summary: [<p>"Der Teuto ruft!"{/* [LInk zu Teuto unten] */} is a community event in Bielefeld where iGEM Bielefeld participated to raise awareness about cystic fibrosis (CF) and gene therapy. They engaged children through interactive experiments, such as creating lung models and simulating mucus to help them understand the challenges faced by CF patients. Adults were informed about their gene therapy project and had meaningful discussions about the implications of CF treatment. Collaborating with other institutions like the life science student initiative btS, the team expanded their outreach. Despite changeable weather, the event was a success in educating the public and improving science communication.</p>],
months: "June"
},
{
@@ -940,7 +940,7 @@ export const timelinedata: Array<TimelineDatenpunkt> = [
insights: [<p>During our time at Achema, we gained valuable insights into life sciences, with a strong focus on LNP production and lab best practices. Our discussions with experts, including Jutta from a life science company, provided us with innovative solutions and cutting-edge techniques. Her insights into current trends helped us better understand how to advance our project.</p>,
<p>Through various expert interactions, we deepened our knowledge of LNP production, learning key manufacturing techniques essential for developing effective therapeutics. These insights allowed us to optimize our production methods, ensuring more efficient processes moving forward.</p>,
<p>Understanding rigorous quality control processes was another key takeaway, enabling us to implement checks that will boost the reliability and safety of our therapeutics. These comprehensive insights have not only supported our current goals but also prepared us for future challenges, equipping us with the tools to navigate the complexities of research and therapeutic development.</p>],
- implementation: [<p>Discussions with industry experts have provided invaluable insights into the production of LNPs[LINK LNPcycle], a crucial aspect of our project. We have identified innovative manufacturing techniques and quality control procedures that we can integrate into our processes. This knowledge not only supports our immediate objectives but also positions us to overcome future challenges in developing effective therapies.</p>],
+ implementation: [<p>Discussions with industry experts have provided invaluable insights into the production of LNPs{/* [LINK LNPcycle] */}, a crucial aspect of our project. We have identified innovative manufacturing techniques and quality control procedures that we can integrate into our processes. This knowledge not only supports our immediate objectives but also positions us to overcome future challenges in developing effective therapies.</p>],
summary: [<p>During discussions with industry experts at Achema 2024, we gained valuable insights that will have a significant impact on our project and provide us with innovative manufacturing techniques and quality control procedures that we are now integrating into our workflows. These insights will help us to optimise our production processes to achieve more efficient and reliable results in therapeutic development. The new knowledge supports our current goals and prepares us for future challenges in the development of effective therapies</p>],
months: "june",
pictureurl_aim:"https://static.igem.wiki/teams/5247/photos/hp/achema1.webp",