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Medientyp:
E-Artikel
Titel:
Cas9/Nickase-induced allelic conversion by homologous chromosome-templated repair in Drosophila somatic cells
Beteiligte:
Roy, Sitara;
Juste, Sara Sanz;
Sneider, Marketta;
Auradkar, Ankush;
Klanseck, Carissa;
Li, Zhiqian;
Julio, Alison Henrique Ferreira;
del Amo, Victor Lopez;
Bier, Ethan;
Guichard, Annabel
Erschienen:
American Association for the Advancement of Science (AAAS), 2022
Erschienen in:
Science Advances, 8 (2022) 26
Sprache:
Englisch
DOI:
10.1126/sciadv.abo0721
ISSN:
2375-2548
Entstehung:
Anmerkungen:
Beschreibung:
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 Drosophila 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 white 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.