Mechanistic and genetic basis of single-strand templated repair at Cas12a-induced DNA breaks in Chlamydomonas reinhardtii

A - Papers appearing in refereed journals

Ferenczi, A., Chew, Y. P., Kroll, E., Von Koppenfels, C., Hudson, A. and Molnar, A. 2021. Mechanistic and genetic basis of single-strand templated repair at Cas12a-induced DNA breaks in Chlamydomonas reinhardtii. Nature Communications. 12 (6751). https://doi.org/0.1038/s41467-021-27004-12NATURECOMMUNICATIONS|(2021)12:6751|https://doi.org/10.1038/s41467-021-27004-1

AuthorsFerenczi, A., Chew, Y. P., Kroll, E., Von Koppenfels, C., Hudson, A. and Molnar, A.
Abstract

Single-stranded oligodeoxynucleotides (ssODNs) are widely used as DNA repair templates inCRISPR/Cas precision genome editing. However, the underlying mechanisms of single-strandtemplated DNA repair (SSTR) are inadequately understood, constraining rational improve-ments to precision editing. Here we study SSTR at CRISPR/Cas12a-induced DNA double-strand breaks (DSBs) in the eukaryotic model green microalgaChlamydomonas reinhardtii.Wedemonstrate that ssODNs physically incorporate into the genome during SSTR at Cas12a-induced DSBs. This process is genetically independent of the Rad51-dependent homologousrecombination and Fanconi anemia pathways, is strongly antagonized by non-homologousend-joining, and is mediated almost entirely by the alternative end-joining enzyme poly-meraseθ. Thesefindings suggest differences in SSTR betweenC. reinhardtiiand animals. Ourwork illustrates the promising potentially ofC. reinhardtiias a model organism for studyingnuclear DNA repair.

Year of Publication2021
JournalNature Communications
Journal citation12 (6751)
Digital Object Identifier (DOI)https://doi.org/0.1038/s41467-021-27004-12NATURECOMMUNICATIONS|(2021)12:6751|https://doi.org/10.1038/s41467-021-27004-1
Open accessPublished as ‘gold’ (paid) open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeBB/V509188/1
BB/R506163/1
Publisher's version
Output statusPublished
Publication dates
Online19 Nov 2021
Publication process dates
Accepted26 Oct 2021
ISSN2041-1723
PublisherNature Publishing Group

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