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DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A

ABSTRACT: DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A

PROVIDER: PRJNA382980 | ENA |

REPOSITORIES: ENA

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Dataset's files

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			Action	DRS
	SRR5448840.fastq.gz	Fastqsanger.gz
	SRR5448831_1.fastq.gz	Fastqsanger.gz
	SRR5448831_2.fastq.gz	Fastqsanger.gz
	SRR5448841.fastq.gz	Fastqsanger.gz
	SRR5448832_1.fastq.gz	Fastqsanger.gz

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DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [WGBS]

Project description:We demonstrate that dCas9-SunTag-DNMT3A dramatically increased CpG methylation at the HOXA5 locus in human embryonic kidney 293T cells (HEK293T). Furthermore, using a single sgRNA, dCas9-SunTag-DNMT3A was able to methylate a 4.5 kb genomic region and repress HOXA5 gene expression. Reduced representation bisulfite sequencing (RRBS) and RNA-seq showed that dCas9-SunTag-DNMT3A methylated regions of interest with minimal impact on the global DNA methylome and transcriptome.

2017-08-22 | GSE97814 | GEO

DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [RRBS]

2017-08-22 | GSE97813 | GEO

DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [RNA-Seq]

2017-08-22 | GSE97815 | GEO

DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [BS-Seq]

Project description:Here, we demonstrate that dCas9-SunTag-DNMT3A dramatically increased CpG methylation at the HOXA5 locus in human embryonic kidney 293T cells (HEK293T). Furthermore, using a single sgRNA, dCas9-SunTag-DNMT3A was able to methylate a 4.5 kb genomic region and repress HOXA5 gene expression. Reduced representation bisulfite sequencing (RRBS) and RNA-seq showed that dCas9-SunTag-DNMT3A methylated regions of interest with minimal impact on the global DNA methylome and transcriptome.

2017-08-22 | GSE97812 | GEO

DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A.

Project description:BackgroundDNA methylation has widespread effects on gene expression during development. However, our ability to assign specific function to regions of DNA methylation is limited by the poor correlation between global patterns of DNA methylation and gene expression.ResultsHere, we utilize nuclease-deactivated Cas9 protein fused to repetitive peptide epitopes (SunTag) recruiting multiple copies of antibody-fused de novo DNA methyltransferase 3A (DNMT3A) (dCas9-SunTag-DNMT3A) to amplify the local DNMT3A concentration to methylate genomic sites of interest. We demonstrate that dCas9-SunTag-DNMT3A dramatically increases CpG methylation at the HOXA5 locus in human embryonic kidney (HEK293T) cells. Furthermore, using a single guide RNA, dCas9-SunTag-DNMT3A is able to methylate a 4.5-kb genomic region and repress HOXA5 gene expression. Reduced representation bisulfite sequencing and RNA-seq show that dCas9-SunTag-DNMT3A methylates regions of interest with minimal impact on the global DNA methylome and transcriptome.ConclusionsThis effective and precise tool enables site-specific manipulation of DNA methylation and may be used to address the relationship between DNA methylation and gene expression.

| S-EPMC5604343 | biostudies-literature

Homo sapiens

Project description:DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [RRBS]

| PRJNA382988 | ENA

Homo sapiens

Project description:DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [WGBS]

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Homo sapiens

Project description:DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [BS-Seq]

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Homo sapiens

Project description:DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [RNA-Seq]