Dispensable Players: N-WASP and WASP Are Not Crucial for Homology-Directed DNA Repair

ABSTRACT: N-WASP and WASP can induce actin polymerization via Arp2/3 and were reported to be crucial for homology directed repair (HDR) of DNA double-strand breaks (DSB). The underlying mechanism was suggested to involve nuclear actin polymerization, but the mechanistic details were debated. Unexpectedly, we show now that neither WASP nor N-WASP are required for HDR during CRISPR-mediated genome editing. Using knock-out and overexpression of N-WASP and WASP in U2OS cells, we did not detect alterations in total gene editing, HDR, or the ratio of HDR to non-homologous end joining (NHEJ) as assessed by different methods. Furthermore, we could not observe co-localization of HA-tagged WASP or N-WASP with DSB. Finally, while the Arp2/3 inhibitor CK-666 and ARPC4 knockdown by siRNA reduced HDR efficiency i

SUBMITTER: Thu Han Le Phan 

PROVIDER: S-SCDT-10_1038-S44319-026-00771-Y | biostudies-other |

REPOSITORIES: biostudies-other