Protospacer modification improves base editing of a canonical splice site variant and recovery of CFTR function in human airway epithelial cells.

ABSTRACT: Canonical splice site variants affecting the 5' GT and 3' AG nucleotides of introns result in severe missplicing and account for about 10% of disease-causing genomic alterations. Treatment of such variants has proven challenging due to the unstable mRNA or protein isoforms that typically result from disruption of these sites. Here, we investigate CRISPR-Cas9-mediated adenine base editing for such variants in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. We validate a CFTR expression minigene (EMG) system for testing base editing designs for two different targets. We then use the EMG system to test non-standard single-guide RNAs with either shortened or lengthened protospacers to correct the most common cystic fibrosis-causing variant in individuals of A

SUBMITTER: Joynt AT 

PROVIDER: S-EPMC10400809 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature