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. 2019 Jun 11;14:4353–4366. doi: 10.2147/IJN.S199104

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© 2019 Hosseini et al.

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Schematic illustration of the Chol-rich lipid-mediated nanoparticle carrying Cas9/sgRNA plasmids.

Notes: Encapsulation of Cas9 and sgRNA plasmids in DOTAP/DOPE/Cholesterol/Chol-PEG and DOTAP/DOPE/Cholesterol/DSPE-PEG liposomes has happened via electrostatic interactions between the positively charged cationic lipid (DOTAP) and the negatively charged pDNA. By the lipoplex formation, transfection was performed by which the complexes entered the cells through endocytosis. Entrapped pDNAs in DOTAP/DOPE/Cholesterol/Chol-PEG escaped from the endosome while the ones in DOTAP/DOPE/Cholesterol/DSPE-PEG were kept in the endosome and degraded by lysosomal nucleases. Released pDNAs were transferred to the nucleus; after transcription and translation, the ribonucleoprotein complex of Cas9/sgRNA was formed and the chromosome was scanned for the target locus to introduce the double-strand cut for editing.