title: Targeted Epigenetic Remodeling of Endogenous Loci by CRISPR/Cas9-Based Transcriptional Activators Directly Converts Fibroblasts to Neuronal Cells.
abstract: Overexpression of exogenous fate-specifying transcription factors can directly reprogram differentiated somatic cells to target cell types. Here, we show that similar reprogramming can also be achieved through the direct activation of endogenous genes using engineered CRISPR/Cas9-based transcriptional activators. We use this approach to induce activation of the endogenous Brn2, Ascl1, and Myt1l genes (BAM factors) to convert mouse embryonic fibroblasts to induced neuronal cells. This direct activation of endogenous genes rapidly remodeled the epigenetic state of the target loci and induced sustained endogenous gene expression during reprogramming. Thus, transcriptional activation and epigenetic remodeling of endogenous master transcription factors are sufficient for conversion between cell types. The rapid and sustained activation of endogenous genes in their native chromatin context by this approach may facilitate reprogramming with transient methods that avoid genomic integration and provides a new strategy for overcoming epigenetic barriers to cell fate specification.
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
authors.forename: Joshua B
authors.initials: JB
authors.lastname: Black
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Adler
authors.affiliation: Ion Channel Research Unit, Duke University Medical Center, Durham, NC 27710, USA; Division of Cardiology, Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Hong-Gang
authors.initials: HG
authors.lastname: Wang
authors.affiliation: Center for Genomic and Computational Biology, Duke University, Durham, NC 27708, USA; University Program in Genetics and Genomics, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Anthony M
authors.initials: AM
authors.lastname: D'Ippolito
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
authors.forename: Hunter A
authors.initials: HA
authors.lastname: Hutchinson
authors.affiliation: Center for Genomic and Computational Biology, Duke University, Durham, NC 27708, USA; Department of Biostatistics and Bioinformatics, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Reddy
authors.affiliation: Ion Channel Research Unit, Duke University Medical Center, Durham, NC 27710, USA; Department of Neurobiology, Duke University Medical Center, Durham, NC 27710, USA; Division of Cardiology, Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Geoffrey S
authors.initials: GS
authors.lastname: Pitt
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
authors.forename: Kam W
authors.initials: KW
authors.lastname: Leong
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA; Center for Genomic and Computational Biology, Duke University, Durham, NC 27708, USA; Department of Orthopaedic Surgery, Duke University Medical Center, Durham, NC 27710, USA. Electronic address: charles.gersbach@duke.edu.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Gersbach
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology; D044127:Epigenesis, Genetic; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D056426:Genetic Loci; D005819:Genetic Markers; D005822:Genetic Vectors / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D015533:Transcriptional Activation
chemical_list: D005819:Genetic Markers; D000094704:RNA, Guide, CRISPR-Cas Systems; D015534:Trans-Activators; D014157:Transcription Factors