title: Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers.
abstract: Technologies that enable targeted manipulation of epigenetic marks could be used to precisely control cell phenotype or interrogate the relationship between the epigenome and transcriptional control. Here we describe a programmable, CRISPR-Cas9-based acetyltransferase consisting of the nuclease-null dCas9 protein fused to the catalytic core of the human acetyltransferase p300. The fusion protein catalyzes acetylation of histone H3 lysine 27 at its target sites, leading to robust transcriptional activation of target genes from promoters and both proximal and distal enhancers. Gene activation by the targeted acetyltransferase was highly specific across the genome. In contrast to previous dCas9-based activators, the acetyltransferase activates genes from enhancer regions and with an individual guide RNA. We also show that the core p300 domain can be fused to other programmable DNA-binding proteins. These results support targeted acetylation as a causal mechanism of transactivation and provide a robust tool for manipulating gene regulation.
authors: 1] Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA. [2] Center for Genomic &Computational Biology, Duke University, Durham, North Carolina, USA.
authors: Isaac B
authors: 0000000230648532
authors: IB
authors: Hilton
authors: 1] Center for Genomic &Computational Biology, Duke University, Durham, North Carolina, USA. [2] University Program in Genetics and Genomics, Duke University Medical Center, Durham, North Carolina, USA.
authors: Anthony M
authors: AM
authors: D'Ippolito
authors: 1] Center for Genomic &Computational Biology, Duke University, Durham, North Carolina, USA. [2] Department of Cell Biology, Duke University Medical Center, Durham, North Carolina, USA.
authors: Christopher M
authors: CM
authors: Vockley
authors: 1] Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA. [2] Center for Genomic &Computational Biology, Duke University, Durham, North Carolina, USA.
authors: Pratiksha I
authors: PI
authors: Thakore
authors: 1] Center for Genomic &Computational Biology, Duke University, Durham, North Carolina, USA. [2] Department of Pediatrics, Division of Medical Genetics, Duke University Medical Center, Durham, North Carolina, USA.
authors: Gregory E
authors: GE
authors: Crawford
authors: 1] Center for Genomic &Computational Biology, Duke University, Durham, North Carolina, USA. [2] Department of Biostatistics &Bioinformatics, Duke University Medical Center, Durham, North Carolina, USA.
authors: Timothy E
authors: 000000027629061X
authors: TE
authors: Reddy
authors: 1] Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA. [2] Center for Genomic &Computational Biology, Duke University, Durham, North Carolina, USA. [3] Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina, USA.
authors: Charles A
authors: CA
authors: Gersbach
mesh_terms: D000123:Acetyltransferases / Q000235:genetics; D064113:CRISPR-Cas Systems / Q000235:genetics*; D050881:E1A-Associated p300 Protein / Q000235:genetics; D004742:Enhancer Elements, Genetic; D057890:Epigenomics / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D011401:Promoter Regions, Genetic; D017393:RNA Editing / Q000235:genetics; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics; D015533:Transcriptional Activation
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D000123:Acetyltransferases; D050881:E1A-Associated p300 Protein; C494360:EP300 protein, human