Structure and functional evaluation of porcine NANOG that is a single-exon gene and has two pseudogenes

Yang, Fan; Zhang, Jinglong; Liu, Yajun; Cheng, De; Wang, Huayan

doi:10.1016/j.biocel.2014.12.009

Cited by 12 publications

(19 citation statements)

References 56 publications

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“…Results showed that addition of OCT4 and SOX2 significantly enhanced NANOG promoter activity, similar to a previous study. 31 However, addition of OTX2 alone significantly reduced NANOG promoter activity (Figure 5e). To investigate the synergistic effect of combining OTX2 with OCT4/SOX2, we applied OTX2 with OCT4 and SOX2.…”

Section: Resultsmentioning

confidence: 92%

“…31 Within the promoter sequence, seven putative OTX2 binding sites and multiple putative OCT4 and SOX2 binding sites were found on the basis of the online JASPAR software (Figure 5f, left). Double enzyme digestions confirmed the accuracy of these recombinant vectors (Figure 5f, right).…”

Section: Resultsmentioning

confidence: 99%

“…The NANOG reporter vector pGL-NANOG and a series of deletion constructs, including pGL-N1, pGL-N2, and pGL-N3, were constructed as described previously. 31 …”

Section: Methodsmentioning

confidence: 99%

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OTX2 impedes self–renewal of porcine iPS cells through downregulation of NANOG expression

Wang

Xie

et al. 2016

Cell Death Discovery

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The transcription factor Otx2 acts as a negative switch in the regulation of transition from naive to primed pluripotency in mouse pluripotent stem cells. However, the molecular features and function of porcine OTX2 have not been well elucidated in porcine-induced pluripotent stem cells (piPSCs). By studying high-throughput transcriptome sequencing and interfering endogenous OTX2 expression, we demonstrate that OTX2 is able to downgrade the self-renewal of piPSCs. OTX2 is highly expressed in porcine brain, reproductive tissues, and preimplantation embryos, but is undetectable in fibroblasts and most somatic tissues. However, the known piPSC lines reported previously produced different levels of OTX2 depending on the induction procedures and culture conditions. Overexpression of porcine OTX2 can reduce the percentage of alkaline phosphatase-positive colonies and downregulate NANOG and OCT4 expression. In contrast, knockdown of OTX2 can significantly increase endogenous expressions of NANOG, OCT4, and ESRRB, and stabilize the pluripotent state of piPSCs. On the other hand, NANOG can directly bind to the OTX2 promoter as shown in ChIP-seq data and repress OTX2 promoter activity in a dose-dependent manner. These observations indicate that OTX2 and NANOG can form a negative feedback circuitry to regulate the pluripotency of porcine iPS cells.

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Section: Resultsmentioning

confidence: 92%

Section: Resultsmentioning

confidence: 99%

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OTX2 impedes self–renewal of porcine iPS cells through downregulation of NANOG expression

Wang

Xie

et al. 2016

Cell Death Discovery

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“…To investigate the regulatory function of OCT4 distal enhancer element, a 676 bp fragment of conserved region 4 (CR4) was subcloned into pGL3‐promoter (E1761, Promega) to make reporter vector pOL676. Nanog reporter constructs used in this study were reported previously (Yang, Zhang, Liu, Cheng, & Wang, ). Primers used to amplify SMAD genes, OCT4 promoter, and other genes are listed in Supplementary Table S1.…”

Section: Methodsmentioning

confidence: 99%

Activin‐SMAD signaling is required for maintenance of porcine iPS cell self‐renewal through upregulation of NANOG and OCT4 expression

Yang

Wang²,

Wang³

et al. 2017

Journal Cellular Physiology

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Porcine induced pluripotent stem cells (piPSCs) retain the enormous potential for farm animal reproduction and translational medicine, and have been reported by many laboratories worldwide. Some piPSC lines were bFGF-dependence and showed mouse EpiSC-like morphology; other lines were LIF-dependence and showed mouse ESC-like morphology. Metastable state of piPSC line that required both LIF and bFGF was also reported. Because bona fide pig embryonic stem cells were not available, uncovering piPSC state-specific regulatory circuitries was the most important task. In this study, we explored the function of Activin-SMAD signaling pathway and its downstream activated target genes in piPSCs. Transcriptome analysis showed that genes involved in Activin-SMAD signaling pathway were evidently activated during porcine somatic cell reprogramming, regardless piPSCs were LIF- or bFGF-dependent. Addition of Activin A and overexpression of SMAD2/3 significantly promoted expressions of porcine NANOG and OCT4, whereas inhibition of Activin-SMAD signaling by SB431542 and SMAD7 reduced NANOG and OCT4 expressions, and induced piPSCs differentiation exiting from pluripotent state. Our data demonstrate that activation of Activin-SMAD signaling pathway by addition of Activin A in culture medium is necessary for maintenance of self-renewal in porcine pluripotent stem cells.

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“…In a comparison and analysis of human, murine, and bovine Nanog promoter sequences, the promoters contained highly conserved regions (Rodda et al, 2005). In primordial germ cells (PGCs), Nanog was required for germ cell development (Yang et al, 2015). Recently, Nanog overexpression promoted the expression of the growth and differentiation factor 3 (GDF3), which is a member of the transforming growth factor beta (TGF-β) family (Park et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Exploration of the exonic variations of the iPSC-related <i>Nanog</i> gene and their effects on phenotypic traits in cattle

Zhang¹,

Pan²,

Lin³

et al. 2016

Arch. Anim. Breed.

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Abstract. Nanog is an important pluripotent transcription regulator transforming somatic cells to induced pluripotent stem cells (iPSCs), and its overexpression leads to a high expression of the growth and differentiation factor 3 (GDF3), which affects animal growth traits. Therefore, the aim of this study was to explore the genetic variations within the Nanog gene and their effects on phenotypic traits in cattle. Six novel exonic single nucleotide polymorphisms (SNPs) were found in six cattle breeds. Seven haplotypes were analyzed: TCAACC (0.260), TCAATA (0.039), TCATCC (0.019), TCGACC (0.506), TCGATA (0.137), TCGTCC (0.036), and CT-GATA (0.003). There were strong linkage disequilibriums of SNP1 and SNP2 in Jiaxian cattle as well as of SNP5 and SNP6 in both Jiaxian cattle and Nanyang cattle. Moreover, SNP3, SNP4, and SNP5 were associated with phenotypes. The individuals with GG genotype at the SNP3 locus or AA genotype at the SNP4 locus showed better body slanting length and chest circumference or body height and hucklebone width in Nanyang cattle. The superiority of the SNP5-C allele regarding body height and cannon circumference was observed in Jiaxian cattle. The combination of SNP3 and SNP4 (GG-AA) had positive effects on body height, body slanting length, and chest circumference. These findings may indicate that Nanog, as a regulator of bovine growth traits, could be a candidate gene for marker-assisted selection (MAS) in breeding and genetics in cattle.

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Structure and functional evaluation of porcine NANOG that is a single-exon gene and has two pseudogenes

Cited by 12 publications

References 56 publications

OTX2 impedes self–renewal of porcine iPS cells through downregulation of NANOG expression

OTX2 impedes self–renewal of porcine iPS cells through downregulation of NANOG expression

Activin‐SMAD signaling is required for maintenance of porcine iPS cell self‐renewal through upregulation of NANOG and OCT4 expression

Exploration of the exonic variations of the iPSC-related <i>Nanog</i> gene and their effects on phenotypic traits in cattle

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Structure and functional evaluation of porcine NANOG that is a single-exon gene and has two pseudogenes

Cited by 12 publications

References 56 publications

OTX2 impedes self–renewal of porcine iPS cells through downregulation of NANOG expression

OTX2 impedes self–renewal of porcine iPS cells through downregulation of NANOG expression

Activin‐SMAD signaling is required for maintenance of porcine iPS cell self‐renewal through upregulation of NANOG and OCT4 expression

Exploration of the exonic variations of the iPSC-related &lt;i&gt;Nanog&lt;/i&gt; gene and their effects on phenotypic traits in cattle

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Exploration of the exonic variations of the iPSC-related <i>Nanog</i> gene and their effects on phenotypic traits in cattle