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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doaj-ffbb1424630d415db375e69bc16fdb702021-02-14T12:33:40ZengNature Publishing GroupScientific Reports2045-23222021-02-011111710.1038/s41598-021-82774-4A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesisYu Fu0Zhenglin Zhu1Geng Meng2Rijun Zhang3Yueping Zhang4Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, College of Veterinary Medicine, China Agricultural UniversitySchool of Life Sciences, Chongqing UniversityLaboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, College of Veterinary Medicine, China Agricultural UniversityLaboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, College of Veterinary Medicine, China Agricultural UniversityLaboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, College of Veterinary Medicine, China Agricultural UniversityAbstract 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.https://doi.org/10.1038/s41598-021-82774-4 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yu Fu Zhenglin Zhu Geng Meng Rijun Zhang Yueping Zhang |
spellingShingle |
Yu Fu Zhenglin Zhu Geng Meng Rijun Zhang Yueping Zhang A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis Scientific Reports |
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 Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2021-02-01 |
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. |
url |
https://doi.org/10.1038/s41598-021-82774-4 |
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