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Cas9/Nickase-induced allelic conversion by homologous chromosome-templated repair in Drosophila somatic cells

Repair of double-strand breaks (DSBs) in somatic cells is primarily accomplished by error-prone nonhomologous end joining and less frequently by precise homology-directed repair preferentially using the sister chromatid as a template. Here, a system performs efficient somatic repair of both DSBs and...

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Bibliographic Details
Published in:Science advances 2022-07, Vol.8 (26), p.eabo0721
Main Authors: Roy, Sitara, Juste, Sara Sanz, Sneider, Marketta, Auradkar, Ankush, Klanseck, Carissa, Li, Zhiqian, Julio, Alison Henrique Ferreira, Lopez del Amo, Victor, Bier, Ethan, Guichard, Annabel
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Language:English
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Summary:Repair of double-strand breaks (DSBs) in somatic cells is primarily accomplished by error-prone nonhomologous end joining and less frequently by precise homology-directed repair preferentially using the sister chromatid as a template. Here, a system performs efficient somatic repair of both DSBs and single-strand breaks (SSBs) using intact sequences from the homologous chromosome in a process we refer to as homologous chromosome-templated repair (HTR). Unexpectedly, HTR-mediated allelic conversion at the locus was more efficient (40 to 65%) in response to SSBs induced by Cas9-derived nickases D10A or H840A than to DSBs induced by fully active Cas9 (20 to 30%). Repair phenotypes elicited by Nickase versus Cas9 differ in both developmental timing (late versus early stages, respectively) and the production of undesired mutagenic events (rare versus frequent). Nickase-mediated HTR represents an efficient and unanticipated mechanism for allelic correction, with far-reaching potential applications in the field of gene editing.
ISSN:2375-2548
2375-2548
DOI:10.1126/sciadv.abo0721