title: Nuclear reprogramming with a non-integrating human RNA virus.
abstract: BACKGROUND
  Advances in the field of stem cells have led to novel avenues for generating induced pluripotent stem cells (iPSCs) from differentiated somatic cells. iPSCs are typically obtained by the introduction of four factors--OCT4, SOX2, KLF4, and cMYC--via integrating vectors. Here, we report the feasibility of a novel reprogramming process based on vectors derived from the non-integrating vaccine strain of measles virus (MV).
  
  
  METHODS
  We produced a one-cycle MV vector by substituting the viral attachment protein gene with the green fluorescent protein (GFP) gene. This vector was further engineered to encode for OCT4 in an additional transcription unit.
  
  
  RESULTS
  After verification of OCT4 expression, we assessed the ability of iPSC reprogramming. The reprogramming vector cocktail with the OCT4-expressing MV vector and SOX2-, KLF4-, and cMYC-expressing lentiviral vectors efficiently transduced human skin fibroblasts and formed iPSC colonies. Reverse transcription-polymerase chain reaction and immunostaining confirmed induction of endogenous pluripotency-associated marker genes, such as SSEA-4, TRA-1-60, and Nanog. Pluripotency of derived clones was confirmed by spontaneous differentiation into three germ layers, teratoma formation, and guided differentiation into beating cardiomyocytes.
  
  
  CONCLUSIONS
  MV vectors can induce efficient nuclear reprogramming. Given the excellent safety record of MV vaccines and the translational capabilities recently developed to produce MV-based vectors now used for cancer clinical trials, our MV vector system provides an RNA-based, non-integrating gene transfer platform for nuclear reprogramming that is amenable for immediate clinical translation.
authors.affiliation: Department of Molecular Medicine, and Virology and Gene Therapy Graduate Track, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN, 55905, USA. driscoll.christopher@mayo.edu.
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Driscoll
authors.affiliation: Department of Molecular Medicine, and Virology and Gene Therapy Graduate Track, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN, 55905, USA. tonne.jason@mayo.edu.
authors.forename: Jason M
authors.initials: JM
authors.lastname: Tonne
authors.affiliation: Department of Molecular Medicine, and Virology and Gene Therapy Graduate Track, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN, 55905, USA. elkhatib.moustafa@mayo.edu.
authors.forename: Moustafa
authors.initials: M
authors.lastname: El Khatib
authors.affiliation: Department of Molecular Medicine, and Virology and Gene Therapy Graduate Track, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN, 55905, USA. cattaneo.roberto@mayo.edu.
authors.forename: Roberto
authors.initials: R
authors.lastname: Cattaneo
authors.affiliation: Department of Molecular Medicine, and Virology and Gene Therapy Graduate Track, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN, 55905, USA. ikeda.yasuhiro@mayo.edu.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Ikeda
authors.affiliation: Department of Molecular Medicine, and Virology and Gene Therapy Graduate Track, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN, 55905, USA. devaux.patricia@mayo.edu.
authors.forename: Patricia
authors.initials: P
authors.lastname: Devaux
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000235:genetics; D015415:Biomarkers; D002460:Cell Line; D065150:Cellular Reprogramming / Q000235:genetics* / Q000502:physiology; D002522:Chlorocebus aethiops; D005347:Fibroblasts / Q000166:cytology; D052816:Foreskin / Q000166:cytology; D057809:HEK293 Cells; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D008459:Measles virus / Q000235:genetics*; D051379:Mice; D016513:Mice, SCID; D032383:Myocytes, Cardiac / Q000166:cytology*; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis / Q000235:genetics*; D011509:Proteoglycans / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D012867:Skin / Q000166:cytology; D055746:Stage-Specific Embryonic Antigens / Q000235:genetics; D014709:Vero Cells
chemical_list: D000954:Antigens, Surface; D015415:Biomarkers; D018398:Homeodomain Proteins; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C489427:MYC protein, human; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D011509:Proteoglycans; D016271:Proto-Oncogene Proteins c-myc; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D055746:Stage-Specific Embryonic Antigens; C107064:TRA-1-60 antigen, human; C066584:stage-specific embryonic antigen-4