305 TRANSPOSON-MEDIATED REPROGRAMMING OF LIVESTOCK SOMATIC CELLS TO INDUCED PLURIPOTENT STEM CELLS

T. R. Talluri; D. Hermann; B. Barg-Kues; K. Debowski; R. Behr; Z. Ivics; V. J. Hall; M. A. Rasmussen; P. Hyttel; H. Niemann; W. A. Kues

doi:10.1071/RDv25n1Ab305

RESEARCH ARTICLE

305 TRANSPOSON-MEDIATED REPROGRAMMING OF LIVESTOCK SOMATIC CELLS TO INDUCED PLURIPOTENT STEM CELLS

T. R. Talluri ^A , D. Hermann ^A , B. Barg-Kues ^A , K. Debowski ^B , R. Behr ^B , Z. Ivics ^C , V. J. Hall ^D , M. A. Rasmussen ^D , P. Hyttel ^D , H. Niemann ^A and W. A. Kues ^A

+ Author Affiliations

- Author Affiliations

^A Institute of Farm Animal Genetics, Friedrich-Loeffler-Institute, Mariensee, Lower Saxony, Germany;

^B German Primate Center, Göttingen, Lower Saxony, Germany;

^C Paul-Ehrlich-Institute, Langen, Germany;

^D University of Copenhagen, Copenhagen, Denmark

Reproduction, Fertility and Development 25(1) 300-300 https://doi.org/10.1071/RDv25n1Ab305
Published: 4 December 2012

Abstract

The elusive nature of embryonic stem cells in livestock makes reprogramming of somatic cells to induced pluripotent stem (iPS) cells a promising approach for targeted genetic modifications. The first attempts to produce iPS cells from livestock species were made using retro- and lentiviral vectors, which are associated with an increased risk of insertional mutagenesis and which are not easily removable after reprogramming. Here, we describe a nonviral method for the derivation of porcine and bovine iPS cells, using Sleeping Beauty (SB) and piggyBac (PB) transposon systems. The transposons encode the murine or primate reprogramming factors OCT4, SOX2, KLF4, MYC, and LIN28, separated by self-cleaving peptide sequences, respectively. In addition, the PB transposon cassette contains a NANOG-cDNA. The SB or PB transposon-reprogrammed porcine iPS cells expressed typical markers of embryonic stem cells (SSEA1, SSEA4, TRA-1-60, and endogenous stemness genes), showed long-term proliferation under feeder-free culture conditions, differentiated into cell types of the 3 germ layers in vitro, and formed teratomas after subcutaneous injection into immune-deficient nude mice. Both transposon systems are currently being tested in bovine fibroblasts. The results are a major step towards the derivation of authentic porcine and bovine iPS cells, in which the transposon transgenes can be eliminated after reprogramming.