Small molecule inhibition of ATM kinase increases CRISPR-Cas9 1-bp insertion frequency.

ABSTRACT: Mutational outcomes following CRISPR-Cas9-nuclease cutting in mammalian cells have recently been shown to be predictable and, in certain cases, skewed toward single genotypes. However, the ability to control these outcomes remains limited, especially for 1-bp insertions, a common and therapeutically relevant class of repair outcomes. Here, through a small molecule screen, we identify the ATM kinase inhibitor KU-60019 as a compound capable of reproducibly increasing the fraction of 1-bp insertions relative to other Cas9 repair outcomes. Small molecule or genetic ATM inhibition increases 1-bp insertion outcome fraction across three human and mouse cell lines, two Cas9 species, and dozens of target sites, although concomitantly reducing the fraction of edited alleles. Notably, KU-60019 increa

SUBMITTER: Bermudez-Cabrera HC 

PROVIDER: S-EPMC8387472 | biostudies-literature | 2021 Aug

REPOSITORIES: biostudies-literature