Gene editing of NCF1 loci is associated with homologous recombination and chromosomal rearrangements.

ABSTRACT: CRISPR-based genome editing of pseudogene-associated disorders, such as p47^phox-deficient chronic granulomatous disease (p47 CGD), is challenged by chromosomal rearrangements due to presence of multiple targets. We report that interactions between highly homologous sequences that are localized on the same chromosome contribute substantially to post-editing chromosomal rearrangements. We successfully employed editing approaches at the NCF1 gene and its pseudogenes, NCF1B and NCF1C, in a human cell line model of p47 CGD and in patient-derived human hematopoietic stem and progenitor cells. Upon genetic engineering, a droplet digital PCR-based method identified cells with altered copy numbers, spanning megabases from the edited loci. We attributed the high aberration frequency to th

SUBMITTER: Raimondi F 

PROVIDER: S-EPMC11464842 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature