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Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo

Genome editing with CRISPR-Cas nucleases has been applied successfully to a wide range of cells and organisms. There is, however, considerable variation in the efficiency of cleavage and outcomes at different genomic targets, even within the same cell type. Some of this variability is likely due to...

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Autores principales: Yarrington, Robert M., Verma, Surbhi, Schwartz, Shaina, Trautman, Jonathan K., Carroll, Dana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6156633/
https://www.ncbi.nlm.nih.gov/pubmed/30201707
http://dx.doi.org/10.1073/pnas.1810062115
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author Yarrington, Robert M.
Verma, Surbhi
Schwartz, Shaina
Trautman, Jonathan K.
Carroll, Dana
author_facet Yarrington, Robert M.
Verma, Surbhi
Schwartz, Shaina
Trautman, Jonathan K.
Carroll, Dana
author_sort Yarrington, Robert M.
collection PubMed
description Genome editing with CRISPR-Cas nucleases has been applied successfully to a wide range of cells and organisms. There is, however, considerable variation in the efficiency of cleavage and outcomes at different genomic targets, even within the same cell type. Some of this variability is likely due to the inherent quality of the interaction between the guide RNA and the target sequence, but some may also reflect the relative accessibility of the target. We investigated the influence of chromatin structure, particularly the presence or absence of nucleosomes, on cleavage by the Streptococcus pyogenes Cas9 protein. At multiple target sequences in two promoters in the yeast genome, we find that Cas9 cleavage is strongly inhibited when the DNA target is within a nucleosome. This inhibition is relieved when nucleosomes are depleted. Remarkably, the same is not true of zinc-finger nucleases (ZFNs), which cleave equally well at nucleosome-occupied and nucleosome-depleted sites. These results have implications for the choice of specific targets for genome editing, both in research and in clinical and other practical applications.
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spelling pubmed-61566332018-09-27 Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo Yarrington, Robert M. Verma, Surbhi Schwartz, Shaina Trautman, Jonathan K. Carroll, Dana Proc Natl Acad Sci U S A Biological Sciences Genome editing with CRISPR-Cas nucleases has been applied successfully to a wide range of cells and organisms. There is, however, considerable variation in the efficiency of cleavage and outcomes at different genomic targets, even within the same cell type. Some of this variability is likely due to the inherent quality of the interaction between the guide RNA and the target sequence, but some may also reflect the relative accessibility of the target. We investigated the influence of chromatin structure, particularly the presence or absence of nucleosomes, on cleavage by the Streptococcus pyogenes Cas9 protein. At multiple target sequences in two promoters in the yeast genome, we find that Cas9 cleavage is strongly inhibited when the DNA target is within a nucleosome. This inhibition is relieved when nucleosomes are depleted. Remarkably, the same is not true of zinc-finger nucleases (ZFNs), which cleave equally well at nucleosome-occupied and nucleosome-depleted sites. These results have implications for the choice of specific targets for genome editing, both in research and in clinical and other practical applications. National Academy of Sciences 2018-09-18 2018-09-10 /pmc/articles/PMC6156633/ /pubmed/30201707 http://dx.doi.org/10.1073/pnas.1810062115 Text en Copyright © 2018 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/ This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Yarrington, Robert M.
Verma, Surbhi
Schwartz, Shaina
Trautman, Jonathan K.
Carroll, Dana
Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo
title Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo
title_full Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo
title_fullStr Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo
title_full_unstemmed Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo
title_short Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo
title_sort nucleosomes inhibit target cleavage by crispr-cas9 in vivo
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6156633/
https://www.ncbi.nlm.nih.gov/pubmed/30201707
http://dx.doi.org/10.1073/pnas.1810062115
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