Details

Autor(en) / Beteiligte

• Huang, Yung-Hsin
• Su, Jianzhong
• Lei, Yong
• Brunetti, Lorenzo
• Gundry, Michael C
• Zhang, Xiaotian
• Jeong, Mira
• Li, Wei
• Goodell, Margaret A

Titel

DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A

Ist Teil von

• Genome Biology, 2017-09, Vol.18 (1), p.176-176, Article 176

Ort / Verlag

England: BioMed Central

Erscheinungsjahr

2017

Quelle

MEDLINE

Beschreibungen/Notizen

• DNA 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. Here, 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. This 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.