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CHIP-seq analysis about Histones and Zscan4 after Zscan4 overexpression

ABSTRACT: CHIP-seq analysis about Histones and Zscan4 after Zscan4 overexpression

PROVIDER: PRJNA515624 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11414081_1.fastq.gz	Fastqsanger.gz
	SRR11414081_2.fastq.gz	Fastqsanger.gz
	SRR11414082_1.fastq.gz	Fastqsanger.gz
	SRR11414082_2.fastq.gz	Fastqsanger.gz
	SRR8450750_1.fastq.gz	Fastqsanger.gz

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CHIP-seq analysis about Histones and Zscan4 after Zscan4 overexpression

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Project description:The gold standard for examining pluripotency of stem cells is to see whether cells can contribute to entire body of animals. Here we show that the increased frequency of Zscan4 activation in mouse ES cells not only enhances, but also maintains their developmental potency in long-term cell culture. As the potency increases, even a whole animal can be produced from a single ES cell injected into 4N blastocyst at unusually high success rate. Although Zscan4-activated cells express genes that are also expressed in 2-cell stage mouse embryos, transiently Zscan4-activated state of ES cells is not associated with the high potency of ES cells. It is thus concluded that ES cells acquire higher potency by going through transient Zscan4 activation state more frequently than the regular state. Taken together, our results indicate that frequent activation of Zscan4 can rejuvenate pluripotent stem cells. Previously, we have shown that constitutive and strong expression of Zscan4 arrest the proliferation of ES cells (refs). We sought for the system to mimic the transient expression of Zscan4 and decided to use ERT2 - Tamoxifen-inducible system (refs). The system allows one to keep a transgene off in the absence of Tmx and turn on in the presence of Tmx at will (refs). We first made a plasmid construct pCAG-Zscan4-ERT2, in which ZSCAN4 ORF fused with ERT2 domain can be driven by a strong ubiquitous promoter CAG. To further investigate this, we transfected pCAG-Zscan4-ERT2 plasmid into an F1 hybrid (C57BL/6J vs. 129) ES cell line V6.5 and isolated multiple cell clones. Based on the qRT-PCR analysis of Zscan4 ORF, we found that cell clones showed a variety of Zscan4 expression levels. We selected the clone that showed the highest Zscan4 expression levels (clone #18) and a clone #2 with the background Zscan4 level.

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