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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 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.

SUBMITTER: Fu Y

PROVIDER: S-EPMC7870972 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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

Fu Yu Y Zhu Zhenglin Z Meng Geng G Zhang Rijun R Zhang Yueping Y

Scientific reports 20210208 1

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 ...[more]

PMID: 33558622

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