title: Non-viral reprogramming of fibroblasts into induced pluripotent stem cells by Sleeping Beauty and piggyBac transposons.
abstract: The generation of induced pluripotent stem (iPS) cells represents a promising approach for innovative cell therapies. The original method requires viral transduction of several reprogramming factors, which may be associated with an increased risk of tumorigenicity. Transposition of reprogramming cassettes represents a recent alternative to viral approaches. Since binary transposons can be produced as common plasmids they provide a safe and cost-efficient alternative to viral delivery methods. Here, we compared the efficiency of two different transposon systems, Sleeping Beauty (SB) and piggyBac (PB), for the generation of murine iPS. Murine fibroblasts derived from an inbred BL/6 mouse line carrying a pluripotency reporter, Oct4-EGFP, and fibroblasts derived from outbred NMRI mice were employed for reprogramming. Both transposon systems resulted in the successful isolation of murine iPS cell lines. The reduction of the core reprogramming factors to omit the proto-oncogene c-Myc was compatible with iPS cell line derivation, albeit with reduced reprogramming efficiencies. The transposon-derived iPS cells featured typical hallmarks of pluripotency, including teratoma growth in immunodeficient mice. Thus SB and PB transposons represent a promising non-viral approach for iPS cell derivation.
authors.affiliation: Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Neustadt, Germany; National Research Center on Equines, Hisar, India.
authors.forename: Thirumala R
authors.initials: TR
authors.lastname: Talluri
authors.affiliation: Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Neustadt, Germany; Central Institute for Research on Buffaloes, Hisar, India.
authors.forename: Dharmendra
authors.initials: D
authors.lastname: Kumar
authors.affiliation: Medical School Hannover, Hannover, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Glage
authors.affiliation: Medical School Hannover, Hannover, Germany.
authors.forename: Wiebke
authors.initials: W
authors.lastname: Garrels
authors.affiliation: Paul-Ehrlich-Institute, Langen, Germany.
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Ivics
authors.affiliation: German Primate Center, Göttingen, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Debowski
authors.affiliation: German Primate Center, Göttingen, Germany.
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Behr
authors.affiliation: Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Neustadt, Germany. Electronic address: wilfried.kues@fli.bund.de.
authors.forename: Wilfried A
authors.initials: WA
authors.lastname: Kues
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002460:Cell Line; D002477:Cells; D004251:DNA Transposable Elements / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology*; D000090063:Proto-Oncogene Mas; D014162:Transfection; D019895:Transposases / Q000235:genetics*; D014780:Viruses / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; C000715470:MAS1 protein, human; D009419:Nerve Tissue Proteins; C569369:PGBD1 protein, human; D000090063:Proto-Oncogene Mas; D019895:Transposases; C494471:sleeping beauty transposase, human
keywords: Genetic background; Inbred; Outbred; Pluripotency reporter; Stemness; Transposition