Targeting the CFTR Locus for Human CFTR Gene Correction using the CRISPR-Cas9 System

Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated nuclease 9 (Cas9), a gene editing tool, is being explored as a possible therapy for CF. Our strategy is to use helper dependent adenoviral (HD-Ad) vectors to deliver the CFTR transgene and the CRISPR-Cas9 system targeting the CFTR locus to human cells. In vitro experiments, using a lacZ donor plasmid, were performed to validate genome targeting efficiency, integration and expression in human embryonic kidney (HEK293) cells. The information gained from these proof-of-concept experiments allowed for the design of HD-Ad vectors. Future experiments using these HD-Ad vectors will allow us to examine CFTR expression and channel activity in CF corrected cells. Overall, these studies will contribute to the development of a long-lasting effective treatment for all CF patients.M.Sc.2021-11-19 00:00:0