title: The piggyBac Transposon as a Platform Technology for Somatic Cell Reprogramming Studies in Mouse.
abstract: Somatic cell reprogramming to induced pluripotent stem cells (iPSCs) is a revolutionary technology, with repercussions affecting modern functional genomics and regenerative medicine. Still, relatively little is known about the processes underlying this dramatic cellular and molecular metamorphosis. Reprogramming technology based on the implementation of piggyBac (PB) transposons has enabled studies of iPSC reprogramming mechanisms, shedding an increasing light on these processes. Unique characteristics of PB transposons such as efficient genomic integration, unlimited cargo capacity, robust gene expression, and even seamless excision highlight the importance of this transgenic tool in advancing stem cell biology. In this chapter, we provide a detailed overview of versatile primary iPSC generation from mouse somatic cells using PB transposons, and the subsequent establishment of robust secondary reprogramming systems. These protocols are highlighted with examples from recent studies as to how PB has been, and continues to be, conducive to the dissection of reprogramming processes at the cellular and molecular levels.
authors.affiliation: Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, 606-8507, Japan. woltjen@cira.kyoto-u.ac.jp.
authors.forename: Knut
authors.initials: K
authors.lastname: Woltjen
authors.affiliation: Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Shin-Il
authors.initials: SI
authors.lastname: Kim
authors.affiliation: Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, M5G 1X5, Canada.
authors.forename: Andras
authors.initials: A
authors.lastname: Nagy
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D058948:Cell Tracking / Q000379:methods; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D002999:Clone Cells / Q000166:cytology; D015925:Cryopreservation; D004251:DNA Transposable Elements / Q000235:genetics*; D004318:Doxycycline / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D000071317:Nanog Homeobox Protein; D000097573:Red Fluorescent Protein
chemical_list: D004251:DNA Transposable Elements; D004318:Doxycycline; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D000071317:Nanog Homeobox Protein; D000097573:Red Fluorescent Protein; C475540:Nanog protein, mouse
keywords: Doxycycline regulation; Induced pluripotent stem cells (iPSCs); Reprogramming; Secondary reprogramming; Transposon; piggyBac