Timed inhibition of CDC7 increases CRISPR-Cas9 mediated templated repair.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 32355159.
- Also identified by DOI 10.1038/s41467-020-15845-1 and PMC identifier 7193628.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Repair of double strand DNA breaks (DSBs) can result in gene disruption or gene modification via homology directed repair (HDR) from donor DNA. Altering cellular responses to DSBs may rebalance editing outcomes towards HDR and away from other repair outcomes. Here, we utilize a pooled CRISPR screen to define host cell involvement in HDR between a Cas9 DSB and a plasmid double stranded donor DNA (dsDonor). We find that the Fanconi Anemia (FA) pathway is required for dsDonor HDR and that other genes act to repress HDR. Small molecule inhibition of one of these repressors, CDC7, by XL413 and other inhibitors increases the efficiency of HDR by up to 3.5 fold in many contexts, including primary T cells. XL413 stimulates HDR during a reversible slowing of S-phase that is unexplored for Cas9-induced HDR. We anticipate that XL413 and other such rationally developed inhibitors will be useful tools for gene modification.
Medical subject headings
- CRISPR-Cas Systems
- Cell Cycle Proteins
- Protein Serine-Threonine Kinases
- Recombinational DNA Repair