Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM)

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  • Kai Xiong, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Grønnegårdsvej 7, 1870 Frederiksberg C, Denmark.
  • ,
  • Yan Zhou
  • ,
  • Poul Hyttel
  • Lars Bolund
  • Kristine Karla Freude, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Grønnegårdsvej 7, 1870 Frederiksberg C, Denmark. Electronic address: kkf@sund.ku.dk.
  • ,
  • Yonglun Luo

Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4 and C-MYC. We have validated that the reprogramming cassette is silenced in the SAM iPSC clones. Expression of pluripotency genes (OCT4, SOX2, LIN28A, NANOG, GDF3, SSEA4, and TRA-1-60), differentiation potential to all three germ layers, and normal karyotypes are validated. These SAM-iPSCs provide a novel, useful tool to investigate genetic regulation of stem cell proliferation and differentiation through CRISPR-mediated activation of endogenous genes.

OriginalsprogEngelsk
TidsskriftStem Cell Research
Vol/bind17
Nummer3
Sider (fra-til)665-669
Antal sider5
ISSN1873-5061
DOI
StatusUdgivet - 17 nov. 2016

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