Cas9 is mostly orthogonal to human systems of DNA break sensing and repair

Cas9 is mostly orthogonal to human systems of DNA break sensing and repair Full article

Journal

PLoS ONE
ISSN: 1932-6203

Output data

Year: 2023, Volume: 18, Number: 11, Article number : e0294683, Pages count : DOI: 10.1371/journal.pone.0294683

Authors

Maltseva Ekaterina A. ¹ , Vasil`eva Inna A. ¹ , Moor Nina A. ¹ , Kim Daria V. ^2,1 , Dyrkheeva Nadezhda S. ¹ , Kutuzov Mikhail M. ¹ , Vokhtantsev Ivan P. ¹ , Kulishova Lilya M. ¹ , Zharkov Dmitry O. ^2,1 , Lavrik Olga I. ^2,1

Affiliations

1	SB RAS Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia
2	Novosibirsk State University, Novosibirsk, Russia

Funding (2)

1	МИНИСТЕРСТВО НАУКИ И ВЫСШЕГО ОБРАЗОВАНИЯ РОССИЙСКОЙ ФЕДЕРАЦИИ	ПФНИ РФ (2021-2030) 0245-2021-0002
2	Russian Science Foundation	РНФ №21-64-00017

CRISPR/Cas9 system is powerful gene editing tool based on the RNA-guided cleavage of target DNA. The Cas9 activity can be modulated by proteins involved in DNA damage signalling and repair due to their interaction with double- and single-strand breaks (DSB and SSB, respectively) generated by wild-type Cas9 or Cas9 nickases. Here we address the interplay between Streptococcus pyogenes Cas9 and key DNA repair factors, including poly (ADP-ribose) polymerase 1 (SSB/DSB sensor), its closest homolog poly(ADP-ribose) polymerase 2, Ku antigen (DSB sensor), DNA ligase I (SSB sensor), replication protein A (DNA duplex destabilizer), and Y-box binding protein 1 (RNA/DNA binding protein). None of those significantly affected Cas9 activity, while Cas9 efficiently shielded DSBs and SSBs from their sensors. Poly(ADP-ribosyl)ation of Cas9 detected for poly(ADP-ribose) polymerase 2 had no apparent effect on the activity. In cellulo, Cas9-dependent gene editing was independent of poly(ADP-ribose) polymerase 1. Thus, Cas9 can be regarded as an enzyme mostly orthogonal to the natural regulation of human systems of DNA break sensing and repair.

Maltseva E.A. , Vasil`eva I.A. , Moor N.A. , Kim D.V. , Dyrkheeva N.S. , Kutuzov M.M. , Vokhtantsev I.P. , Kulishova L.M. , Zharkov D.O. , Lavrik O.I.
Cas9 is mostly orthogonal to human systems of DNA break sensing and repair
PLoS ONE. 2023. V.18. N11. e0294683 . DOI: 10.1371/journal.pone.0294683 WOS Scopus PMID OpenAlex

Published print:

Nov 29, 2023

Web of science:	WOS:001142779200209
Scopus:	2-s2.0-85178223768
PMID:	38019812
OpenAlex:	W4389128832

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