title: Enhanced MyoD-induced transdifferentiation to a myogenic lineage by fusion to a potent transactivation domain.
abstract: Genetic reprogramming holds great potential for disease modeling, drug screening, and regenerative medicine. Genetic reprogramming of mammalian cells is typically achieved by forced expression of natural transcription factors that control master gene networks and cell lineage specification. However, in many instances, the natural transcription factors do not induce a sufficiently robust response to completely reprogram cell phenotype. In this study, we demonstrate that protein engineering of the master transcription factor MyoD can enhance the conversion of human dermal fibroblasts and adult stem cells to a skeletal myocyte phenotype. Fusion of potent transcriptional activation domains to MyoD led to increased myogenic gene expression, myofiber formation, cell fusion, and global reprogramming of the myogenic gene network. This work supports a general strategy for synthetically enhancing the direct conversion between cell types that can be applied in both synthetic biology and regenerative medicine.
authors.forename: Ami M
authors.initials: AM
authors.lastname: Kabadi
authors.forename: Pratiksha I
authors.initials: PI
authors.lastname: Thakore
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Vockley
authors.forename: David G
authors.initials: DG
authors.lastname: Ousterout
authors.forename: Tyler M
authors.initials: TM
authors.lastname: Gibson
authors.affiliation: §Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina 27710, United States.
authors.forename: Farshid
authors.initials: F
authors.lastname: Guilak
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Reddy
authors.affiliation: §Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina 27710, United States.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Gersbach
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism; D000595:Amino Acid Sequence; D019070:Cell Lineage; D054338:Cell Transdifferentiation; D065150:Cellular Reprogramming; D020405:Dermis / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D005822:Genetic Vectors / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008969:Molecular Sequence Data; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism; D017570:MyoD Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering; D017434:Protein Structure, Tertiary
chemical_list: D017570:MyoD Protein
keywords: MyoD; adipose stem cell; genetic reprogramming; mesenchymal stem cell; muscle cell therapy; myogenesis; protein engineering; regenerative medicine; transcription factor