title: A CRISPR/Cas9-based system for reprogramming cell lineage specification.
abstract: Gene activation by the CRISPR/Cas9 system has the potential to enable new approaches to science and medicine, but the technology must be enhanced to robustly control cell behavior. We show that the fusion of two transactivation domains to Cas9 dramatically enhances gene activation to a level that is necessary to reprogram cell phenotype. Targeted activation of the endogenous Myod1 gene locus with this system led to stable and sustained reprogramming of mouse embryonic fibroblasts into skeletal myocytes. The levels of myogenic marker expression obtained by the activation of endogenous Myod1 gene were comparable to that achieved by overexpression of lentivirally delivered MYOD1 transcription factor.
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA; Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA.
authors.forename: Syandan
authors.initials: S
authors.lastname: Chakraborty
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA; Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA.
authors.forename: HaYeun
authors.initials: H
authors.lastname: Ji
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
authors.forename: Ami M
authors.initials: AM
authors.lastname: Kabadi
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA; Institute for Genome Sciences and Policy, Duke University, Durham, NC 27708, USA; Department of Orthopaedic Surgery, Duke University Medical Center, Durham, NC 27708, USA.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Gersbach
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA; Department of Biomedical Engineering, Khalifa University, Abu Dhabi 127788, UAE.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Christoforou
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA; Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA. Electronic address: kam.leong@duke.edu.
authors.forename: Kam W
authors.initials: KW
authors.lastname: Leong
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics*; D065150:Cellular Reprogramming; D015870:Gene Expression; D005786:Gene Expression Regulation; D051379:Mice; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism; D009928:Organ Specificity / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D011993:Recombinant Fusion Proteins