<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 [1]. 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 <SupScrollLinklabel="1"></SupScrollLink>. 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>
<H5text="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>
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@@ -542,7 +544,7 @@ export function Results() {
/>
</div>
<divclassName='col'>
<p>In our first set of experiments, we measured current density in <aonClick={()=>goToPageAndScroll ('Cell Culture','/materials-methods')}>HEK293T CFTR wild type (WT) and HEK293T F508del</a> cell lines, comparing them with regular HEK293. The results demonstrated significant differences in chloride ion conductance, with the CFTR-expressing cell lines showing enhanced conductivity compared to HEK293 (Figure 37). However, a drawback was that we did not observe any significant differences between the HEK293T CFTR WT and F508del cell line. This was unexpected, as the F508del mutation typically leads to a knockdown of the CFTR protein[2], impairing chloride ion transport through the CFTR channel.</p>
<p>In our first set of experiments, we measured current density in <aonClick={()=>goToPageAndScroll ('Cell Culture','/materials-methods')}>HEK293T CFTR wild type (WT) and HEK293T F508del</a> cell lines, comparing them with regular HEK293. The results demonstrated significant differences in chloride ion conductance, with the CFTR-expressing cell lines showing enhanced conductivity compared to HEK293 (Figure 37). However, a drawback was that we did not observe any significant differences between the HEK293T CFTR WT and F508del cell line. This was unexpected, as the F508del mutation typically leads to a knockdown of the CFTR protein<SupScrollLinklabel="2"></SupScrollLink>, impairing chloride ion transport through the CFTR channel.</p>
</div>
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<H4text="Further Validation and Challenges"/>
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@@ -568,6 +570,7 @@ export function Results() {
<p><DownloadLinkurl="https://static.igem.wiki/teams/5247/pdfs/raw-data-patch-clamp.pdf"fileName="raw-data-patch-clamp.pdf"/> Supplementary Material for Patch Clamp</p>
<DownloadLinkurl="https://static.igem.wiki/teams/5247/pdfs/raw-data-delivery.pdf"fileName="raw-data-delivery.pdf"/> Supplementary Material for Delivery
author = {Wang, X. and Liu, S. and Sun, Y. and others},
title = {Preparation of selective organ-targeting (SORT) lipid nanoparticles (LNPs) using multiple technical methods for tissue-specific mRNA delivery},
journal = {Nature Protocols},
volume = {18},
pages = {265--291},
year = {2023},
doi = {10.1038/s41596-022-00755-},
url = {https://doi.org/10.1038/s41596-022-00755-}
}
`,
`@article{sousa2024primeediting,
author = {Sousa, A. A. and Hemez, C. and Lei, L. and others},
title = {Systematic optimization of prime editing for the efficient functional correction of CFTR F508del in human airway epithelial cells},