<p>By focusing on the F508del mutation, we also hope to contribute valuable insights to the global Cystic Fibrosis community. Although this mutation is most common in European populations, it is also found in other regions around the world<SupScrollLinklabel="4"/><sup>,</sup><SupScrollLinklabel="5"/>. Our research could thus help inform treatment strategies and health policies on an international scale. </p>
<p>By focusing on the F508del mutation, we also hope to contribute valuable insights to the global Cystic Fibrosis community. Although this mutation is most common in European populations, it is also found in other regions around the world<SupScrollLinklabel="5"/>. Our research could thus help inform treatment strategies and health policies on an international scale. </p>
<p>With several team members focusing their studies on biomedical fields, we began by examining the current landscape of CF treatments. It quickly became clear that, despite recent progress, there is still no cure. Most therapies, such as CFTR modulators, focus on managing symptoms and improving lung function rather than addressing the underlying cause of the disease <SupScrollLinklabel="6"/> . This realization led us to explore gene-editing technologies, thus leading us to Prime Editing—a next generation gene editing method—captured our attention. </p>
<p>While Prime Editing holds great promise, we found that its application for Cystic Fibrosis, particularly the F508del mutation, had not been fully explored. Recognizing this gap in the research inspired us to take on the challenge of optimizing Prime Editing for this specific mutation. Our mission became clear: we want to contribute to the development of a potential therapeutic approach for Cystic Fibrosis, specifically targeting the F508del mutation with prime editing, and bring us closer to a long-term solution for patients. </p>
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<p>Overall, there are many different Prime Editing systems with a variety of components and complexity, starting from PE2 up to PE7. Possible edits could integrate substitutions, inserts and deletions in the range of one base up to hundreds of nucleotides, with gradually decreasing editing efficiency. Therefore Prime Editing technology allows targeted modifications of specific genes. </p>
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<p>However, the Prime Editing complex is relatively large, posing challenges for therapeutic delivery. Additionally, Prime Editing has been shown to be relatively inefficient in terms of gene editing rates, which could limit its therapeutic utility<SupScrollLinklabel="4"/>. Our project aims to enhance the Prime Editing approach by miniaturizing its components and enhancing its efficiency, as well as precision. </p>
<p>However, the Prime Editing complex is relatively large, posing challenges for therapeutic delivery. Additionally, Prime Editing has been shown to be relatively inefficient in terms of gene editing rates, which could limit its therapeutic utility. Our project aims to enhance the Prime Editing approach by miniaturizing its components and enhancing its efficiency, as well as precision. </p>
<p>As shown in the image, we developed two potential configurations for Prime Guide, each using a different nickase: one based on the Fanzor (nSpuFz1) nickase and the other on a CasX (nPlmCasX) nickase. Both configurations are designed to improve the precision and stability of the Prime Editing system. The pegRNA scaffold, reverse transcriptase (PE6c), and primer binding site (PBS/RTT) work together in both systems to introduce precise edits, with the La(1-193) enhancing stability and function.</p>
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description={<span> Endosomal escape vs degradation of LNP cargo at endocytosis<SupScrollLinklabel="4"/>.</span>}
description={<span> Endosomal escape vs degradation of LNP cargo at endocytosis.</span>}
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<p>LNPs are pivotal not only for shielding mRNA but also for ensuring its efficient delivery into target cells. They facilitate cellular uptake through endocytosis, where the cell membrane engulfs the nanoparticle. LNPs are acclaimed for their high drug-loading capacities, which greatly enhance their therapeutic effectiveness. However, the success of this delivery hinges on effective endosomal escape. Ideally, LNPs release their mRNA payload into the cytoplasm after escaping from endosomes. If this escape process is inefficient, the mRNA can be degraded by lysosomes, which poses a significant challenge for mRNA vaccines and therapies<SupScrollLinklabel="4"/>.</p>
<p>LNPs are pivotal not only for shielding mRNA but also for ensuring its efficient delivery into target cells. They facilitate cellular uptake through endocytosis, where the cell membrane engulfs the nanoparticle. LNPs are acclaimed for their high drug-loading capacities, which greatly enhance their therapeutic effectiveness. However, the success of this delivery hinges on effective endosomal escape. Ideally, LNPs release their mRNA payload into the cytoplasm after escaping from endosomes. If this escape process is inefficient, the mRNA can be degraded by lysosomes, which poses a significant challenge for mRNA vaccines and therapies.</p>
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<p>A crucial advancement in LNP technology involves the use of pH-sensitive cationizable lipids. These lipids remain neutral at physiological pH but become cationic in the acidic environment of endosomes. This shift in charge helps dissociate the nanoparticles and disrupt the endosomal membrane, enhancing the likelihood of successful endosomal escape <SupScrollLinklabel="5"/> . </p>