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

CRISPR/Cas9 system is а 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-...

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Autores principales: Maltseva, Ekaterina A., Vasil’eva, Inna A., Moor, Nina A., Kim, Daria V., Dyrkheeva, Nadezhda S., Kutuzov, Mikhail M., Vokhtantsev, Ivan P., Kulishova, Lilya M., Zharkov, Dmitry O., Lavrik, Olga I.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686484/
https://www.ncbi.nlm.nih.gov/pubmed/38019812
http://dx.doi.org/10.1371/journal.pone.0294683
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author Maltseva, Ekaterina A.
Vasil’eva, Inna A.
Moor, Nina A.
Kim, Daria V.
Dyrkheeva, Nadezhda S.
Kutuzov, Mikhail M.
Vokhtantsev, Ivan P.
Kulishova, Lilya M.
Zharkov, Dmitry O.
Lavrik, Olga I.
author_facet Maltseva, Ekaterina A.
Vasil’eva, Inna A.
Moor, Nina A.
Kim, Daria V.
Dyrkheeva, Nadezhda S.
Kutuzov, Mikhail M.
Vokhtantsev, Ivan P.
Kulishova, Lilya M.
Zharkov, Dmitry O.
Lavrik, Olga I.
author_sort Maltseva, Ekaterina A.
collection PubMed
description CRISPR/Cas9 system is а 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.
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spelling pubmed-106864842023-11-30 Cas9 is mostly orthogonal to human systems of DNA break sensing and repair Maltseva, Ekaterina A. Vasil’eva, Inna A. Moor, Nina A. Kim, Daria V. Dyrkheeva, Nadezhda S. Kutuzov, Mikhail M. Vokhtantsev, Ivan P. Kulishova, Lilya M. Zharkov, Dmitry O. Lavrik, Olga I. PLoS One Research Article CRISPR/Cas9 system is а 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. Public Library of Science 2023-11-29 /pmc/articles/PMC10686484/ /pubmed/38019812 http://dx.doi.org/10.1371/journal.pone.0294683 Text en © 2023 Maltseva et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Maltseva, Ekaterina A.
Vasil’eva, Inna A.
Moor, Nina A.
Kim, Daria V.
Dyrkheeva, Nadezhda S.
Kutuzov, Mikhail M.
Vokhtantsev, Ivan P.
Kulishova, Lilya M.
Zharkov, Dmitry O.
Lavrik, Olga I.
Cas9 is mostly orthogonal to human systems of DNA break sensing and repair
title Cas9 is mostly orthogonal to human systems of DNA break sensing and repair
title_full Cas9 is mostly orthogonal to human systems of DNA break sensing and repair
title_fullStr Cas9 is mostly orthogonal to human systems of DNA break sensing and repair
title_full_unstemmed Cas9 is mostly orthogonal to human systems of DNA break sensing and repair
title_short Cas9 is mostly orthogonal to human systems of DNA break sensing and repair
title_sort cas9 is mostly orthogonal to human systems of dna break sensing and repair
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686484/
https://www.ncbi.nlm.nih.gov/pubmed/38019812
http://dx.doi.org/10.1371/journal.pone.0294683
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