Use of adenine base editing and homology-independent targeted integration strategies to correct the cystic fibrosis causing variant, W1282X.

ABSTRACT: Small molecule drugs known as modulators can treat ~ 90% of people with cystic fibrosis (CF), but do not work for premature termination codon variants such as W1282X (c.3846G > A). Here we evaluated two gene editing strategies, Adenine Base Editing (ABE) to correct W1282X, and Homology-Independent Targeted Integration (HITI) of a CFTR superexon comprising exons 23-27 (SE23-27) to enable expression of a CFTR mRNA without W1282X. In Flp-In-293 cells stably expressing a CFTR expression minigene bearing W1282X, ABE corrected 24% of W1282X alleles, rescued CFTR mRNA from nonsense mediated decay and restored protein expression. However, bystander editing at the adjacent adenine (c.3847A > G), caused an amino acid change (R1283G) that affects CFTR maturation and ablates ion channel activity. In p

SUBMITTER: Mention K 

PROVIDER: S-EPMC10656707 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature