title: Efficient generation of human iPSCs by a synthetic self-replicative RNA.
abstract: The generation of human induced pluripotent stem cells (iPSCs) holds great promise for the development of regenerative medicine therapies to treat a wide range of human diseases. However, the generation of iPSCs in the absence of integrative DNA vectors remains problematic. Here, we report a simple, highly reproducible RNA-based iPSC generation approach that utilizes a single, synthetic self-replicating VEE-RF RNA replicon that expresses four reprogramming factors (OCT4, KLF4, and SOX2, with c-MYC or GLIS1) at consistent high levels prior to regulated RNA degradation. A single VEE-RF RNA transfection into newborn or adult human fibroblasts resulted in efficient generation of iPSCs with all the hallmarks of stem cells, including cell surface markers, global gene expression profiles, and in vivo pluripotency, to differentiate into all three germ layers. The VEE-RF RNA-based approach has broad applicability for the generation of iPSCs for ultimate use in human stem cell therapies in regenerative medicine.
authors.affiliation: Department of Cellular & Molecular Medicine, University of California, San Diego School of Medicine, 9500 Gilman Drive, La Jolla, CA 92093-0686, USA.
authors.forename: Naohisa
authors.initials: N
authors.lastname: Yoshioka
authors.forename: Edwige
authors.initials: E
authors.lastname: Gros
authors.forename: Hai-Ri
authors.initials: HR
authors.lastname: Li
authors.forename: Shantanu
authors.initials: S
authors.lastname: Kumar
authors.forename: Dekker C
authors.initials: DC
authors.lastname: Deacon
authors.forename: Cornelia
authors.initials: C
authors.lastname: Maron
authors.forename: Alysson R
authors.initials: AR
authors.lastname: Muotri
authors.forename: Neil C
authors.initials: NC
authors.lastname: Chi
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Fu
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Yu
authors.forename: Steven F
authors.initials: SF
authors.lastname: Dowdy
mesh_terms: D000328:Adult; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D065150:Cellular Reprogramming; D002999:Clone Cells; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D008297:Male; D051379:Mice; D008819:Mice, Nude; D012313:RNA / Q000378:metabolism*; D012093:Replicon / Q000235:genetics*; D014162:Transfection
chemical_list: C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D012313:RNA
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