Temporal repression of endogenous pluripotency genes during reprogramming of porcine induced pluripotent stem cells

Cellular Reprogramming
Vanessa J HallPoul Hyttel

Abstract

Porcine induced pluripotent stem cells (piPSCs) have the capacity to differentiate in vitro and in vivo and form chimeras. However, the lack of transgene silencing of exogenous DNA integrated into the genome and the inability of cells to proliferate in the absence of transgene expression are underlying reported problems, suggesting that reprogramming is not complete. The aim of the present study was to evaluate reprogramming events using a partially reprogrammed piPSC-like line expressing hOCT4, hNANOG, and hcMYC under tetracycline-regulated control to investigate the effects of these particular transgenes on the expression of the porcine endogenous pluripotency machinery. Endogenous and exogenous gene expression of OCT4, NANOG, SOX2, KLF4, and cMYC was determined at passages 5, 10, 15, and 20, both in cells cultured at 1 μg/mL doxycycline or 4 μg/mL doxycycline. Our results revealed that endogenous genes are repressed by their transgene counterparts in culture and that lack of expression of the transgenes, SOX2 and KLF4 allows for expression of endogenous SOX2 and KLF4. Furthermore, we report that alternate endogenous transcripts for pNANOG, pSOX2, and pKLF4 can also be detected in the pig. Despite the ability for some endogen...Continue Reading

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Citations

Dec 17, 2014·Cellular Reprogramming·Sergio D GermanRamiro Alberio
Dec 19, 2012·Reproduction, Fertility, and Development·Monika Nowak-Imialek, Heiner Niemann
Dec 7, 2013·Reproduction, Fertility, and Development·Ki-Eun Park, Bhanu Prakash V L Telugu
Feb 20, 2016·Transgenic Research·Delia A Soto, Pablo J Ross
Nov 14, 2015·Annual Review of Animal Biosciences·Toshihiko EzashiR Michael Roberts
Jan 1, 2019·Reproduction, Fertility, and Development·Micaela NavarroPablo J Ross
Feb 24, 2016·Journal of Animal Science and Biotechnology·J OgorevcP Dovč
Oct 25, 2020·Journal of Animal Science·Yue SuYoung Tang
Dec 10, 2020·BMC Veterinary Research·Rachel A ScarfoneThomas G Koch

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Methods Mentioned

BETA
PCR
antisense oligonucleotides
transmission electron microscopy

Software Mentioned

Addgene
Leica Application suite

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