RRC ID 61088
Author Hanasaki M, Masumoto H.
Title CRISPR/Transposon gene integration (CRITGI) can manage gene expression in a retrotransposon-dependent manner.
Journal Sci Rep
Abstract The fine-tuning of gene expression contributes to both basic science and applications. Here, we develop a novel gene expression technology termed CRITGI (CRISPR/Transposon gene integration). CRITGI uses CRISPR/Cas9 to integrate multiple copies of the plasmid pTy1 into Ty1 loci, budding yeast retrotransposons. The pTy1 plasmid harbors a Ty1 consensus sequence for integration, a gene of interest with its own promoter and a selection marker gene. Interestingly, the expression of the pTy1 gene in Ty1 loci could be induced in synthetic complete amino acid depletion medium, which could activate the selection marker gene on pTy1. The induction or repression of the gene on pTy1 depended on Ty1 transcription. Activation of the selection marker gene on pTy1 triggered Ty1 transcription, which led to induction of the gene on pTy1. The gene on pTy1 was not transcribed with Ty1 mRNA; the transcription required its own promoter. Furthermore, the trimethylation of histone H3 on lysine 4, a landmark of transcriptionally active chromatin, accumulated at the 5' end of the gene on pTy1 following selection marker gene activation. Thus, CRITGI is a unique gene regulation system to induce the genes on pTy1 in amino acid depletion medium and utilizes Ty1 transcription to create a chromatin environment favorable for the transcription of the genes on pTy1.
Volume 9(1)
Pages 15300
Published 2019-10-25
DOI 10.1038/s41598-019-51891-6
PII 10.1038/s41598-019-51891-6
PMID 31653950
PMC PMC6814769
MeSH Binding Sites / genetics CRISPR-Cas Systems* Chromatin / genetics Chromatin / metabolism Gene Expression Regulation, Fungal* Genes, Fungal / genetics Histones / metabolism Lysine / metabolism Methylation Models, Genetic Mutagenesis, Insertional / genetics Promoter Regions, Genetic / genetics Retroelements / genetics* Saccharomyces cerevisiae / genetics* Saccharomyces cerevisiae / metabolism Saccharomyces cerevisiae Proteins / genetics* Saccharomyces cerevisiae Proteins / metabolism Transcription, Genetic Transcriptional Activation
IF 3.998
DNA material PHM661 (RDB18135) PHM688 (RDB18136) PHM663 (RDB18137) PHM664 (RDB18138) PHM740 (RDB18139) PHM760 (RDB18140) PHM745 (RDB18141) PHM757 (RDB18142) PHM829 (RDB18143) PHM820 (RDB18144) PHM833 (RDB18145) PHM821 (RDB18146) PHM802 (RDB18147)