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Question

Representative sequencing result showed the successful HDR correction event at G542X mutation site was observed in mouse lungs after DOTAP40-HDR, by subsequent cloning and DNA sequencing

CRISPR/Cas9 was used to improve gene correction at the G542X mutation site in lung epithelial cells, potentially offering a promising solution for treating cystic fibrosis. This result is promising, as there are few dividing cell populations that can be corrected through HDR-mediated strategy. This requirement of CRISPR/Cas9 HDR is likely a limiting factor here, which may be overcome in future studies using next-generation gene editors that do not require double-strand breaks. The cloning result further confirmed that successful gene correction at G542X mutation site was observed with DOTAP40-HDR treatment (Fig. 4e).

Answer 1

Representative sequencing results showed successful HDR correction at the G542X mutation site in mouse lungs after DOTAP40-HDR. CRISPR/Cas9 was used to improve gene correction in lung epithelial cells, offering a potential treatment for cystic fibrosis. The result is promising, as few dividing cell populations can be corrected through HDR. The cloning result confirmed successful gene correction at the G542X mutation site with DOTAP40-HDR treatment.