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

Aron Ferenczi, Yen Peng Chew, Erika Kroll, Charlotte Koppenfels, Andrew Hudson and Attila Molnar ()
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Aron Ferenczi: University of Edinburgh
Yen Peng Chew: University of Edinburgh
Erika Kroll: University of Edinburgh
Charlotte Koppenfels: University of Edinburgh
Andrew Hudson: University of Edinburgh
Attila Molnar: University of Edinburgh

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract Single-stranded oligodeoxynucleotides (ssODNs) are widely used as DNA repair templates in CRISPR/Cas precision genome editing. However, the underlying mechanisms of single-strand templated DNA repair (SSTR) are inadequately understood, constraining rational improvements to precision editing. Here we study SSTR at CRISPR/Cas12a-induced DNA double-strand breaks (DSBs) in the eukaryotic model green microalga Chlamydomonas reinhardtii. We demonstrate that ssODNs physically incorporate into the genome during SSTR at Cas12a-induced DSBs. This process is genetically independent of the Rad51-dependent homologous recombination and Fanconi anemia pathways, is strongly antagonized by non-homologous end-joining, and is mediated almost entirely by the alternative end-joining enzyme polymerase θ. These findings suggest differences in SSTR between C. reinhardtii and animals. Our work illustrates the promising potentially of C. reinhardtii as a model organism for studying nuclear DNA repair.

Date: 2021
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-27004-1

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DOI: 10.1038/s41467-021-27004-1

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