A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis

Abstract Post-translational modifications of histone proteins greatly impact gene expression and cell fate decisions in eukaryotes. To study these, it is important to develop a convenient, multiplex, and efficient method to precisely introduce mutations to histones. Because eukaryotic cells usually...

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Autores principales:	Yu Fu, Zhenglin Zhu, Geng Meng, Rijun Zhang, Yueping Zhang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ffbb1424630d415db375e69bc16fdb70
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spelling	oai:doaj.org-article:ffbb1424630d415db375e69bc16fdb702021-12-02T14:11:30ZA CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis10.1038/s41598-021-82774-42045-2322https://doaj.org/article/ffbb1424630d415db375e69bc16fdb702021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82774-4https://doaj.org/toc/2045-2322Abstract Post-translational modifications of histone proteins greatly impact gene expression and cell fate decisions in eukaryotes. To study these, it is important to develop a convenient, multiplex, and efficient method to precisely introduce mutations to histones. Because eukaryotic cells usually contain multiple copies of histone genes, it is a challenge to mutate all histones at the same time by the traditional homologous recombination method. Here, we developed a CRISPR-Cas9 based shuffle system in Saccharomyces cerevisiae, to generate point mutations on both endogenous histone H3 and H4 genes in a rapid, seamless and multiplex fashion. Using this method, we generated yeast strains containing histone triple H3–K4R–K36R–K79R mutants and histone combinatorial H3–K56Q–H4–K59A double mutants with high efficiencies (70–80%). This CRISPR-Cas9 based mutagenesis system could be an invaluable tool to the epigenetics field.Yu FuZhenglin ZhuGeng MengRijun ZhangYueping ZhangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-7 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yu Fu Zhenglin Zhu Geng Meng Rijun Zhang Yueping Zhang A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis
description	Abstract Post-translational modifications of histone proteins greatly impact gene expression and cell fate decisions in eukaryotes. To study these, it is important to develop a convenient, multiplex, and efficient method to precisely introduce mutations to histones. Because eukaryotic cells usually contain multiple copies of histone genes, it is a challenge to mutate all histones at the same time by the traditional homologous recombination method. Here, we developed a CRISPR-Cas9 based shuffle system in Saccharomyces cerevisiae, to generate point mutations on both endogenous histone H3 and H4 genes in a rapid, seamless and multiplex fashion. Using this method, we generated yeast strains containing histone triple H3–K4R–K36R–K79R mutants and histone combinatorial H3–K56Q–H4–K59A double mutants with high efficiencies (70–80%). This CRISPR-Cas9 based mutagenesis system could be an invaluable tool to the epigenetics field.
format	article
author	Yu Fu Zhenglin Zhu Geng Meng Rijun Zhang Yueping Zhang
author_facet	Yu Fu Zhenglin Zhu Geng Meng Rijun Zhang Yueping Zhang
author_sort	Yu Fu
title	A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis
title_short	A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis
title_full	A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis
title_fullStr	A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis
title_full_unstemmed	A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis
title_sort	crispr-cas9 based shuffle system for endogenous histone h3 and h4 combinatorial mutagenesis
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/ffbb1424630d415db375e69bc16fdb70
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A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis

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