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Targeted Epigenetic Remodeling of Endogenous Loci by CRISPR/Cas9-Based Transcriptional Activators Directly Converts Fibroblasts to Neuronal Cells.

TitleTargeted Epigenetic Remodeling of Endogenous Loci by CRISPR/Cas9-Based Transcriptional Activators Directly Converts Fibroblasts to Neuronal Cells.
Publication TypeJournal Article
Year of Publication2016
AuthorsBlack JB, Adler AF, Wang H-G, D'Ippolito AM, Hutchinson HA, Reddy TE, Pitt GS, Leong KW, Gersbach CA
JournalCell Stem Cell
Volume19
Issue3
Pagination406-14
Date Published2016 Sep 01
ISSN1875-9777
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.

DOI10.1016/j.stem.2016.07.001
Alternate JournalCell Stem Cell
PubMed ID27524438
PubMed Central IDPMC5010447
Grant ListT32 GM008555 / GM / NIGMS NIH HHS / United States
DP2 OD008586 / OD / NIH HHS / United States
T32 GM007754 / GM / NIGMS NIH HHS / United States
R01 DA036865 / DA / NIDA NIH HHS / United States
U01 HG007900 / HG / NHGRI NIH HHS / United States
P30 AR066527 / AR / NIAMS NIH HHS / United States
F31 AI124563 / AI / NIAID NIH HHS / United States