Expression analysis of the endogenous Zscan4 locus and its coding proteins in mouse ES cells and preimplantation embryos

Kei ichiro Ishiguro, Yuki Nakatake, Nana Chikazawa-Nohtomi, Hiromi Kimura, Tomohiko Akiyama, Mayumi Oda, Shigeru Ko, Minoru Ko

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Mouse Zinc finger and SCAN domain containing 4 (Zscan4) is encoded in multiple copies of Zscan4 genes, which are expressed in late two-cell stage preimplantation embryos and in 1–5% of the embryonic stem (ES) cell population at a given time. Due to the highly identical nucleotide sequences of multiple copies of Zscan4 paralogs and pseudogenes in the mouse Zscan4 genomic cluster, previous analyses have been done using exogenous transgenes under the regulation of Zscan4c promoter. In this manuscript, we generated knock-in mouse ES cell lines and mouse lines, in which the expression of endogenous Zscan4c, one of the Zscan4 genes, can be specifically monitored with a green fluorescent protein variant, Emerald. Interestingly, we found that only ∼30% of Zscan4-immunopositive ES cells were Emerald positive, suggesting that even when the Zscan4 locus is active, not all Zscan4 genes are expressed synchronously. We also carried out mass spectrometry of protein complexes associated with endogenous Zscan4 proteins. Taken together, our genetic engineering at an endogenous Zscan4c gene provides the first clue for the expression and function of each gene copy of Zscan4 locus in a physiological context.

Original languageEnglish
Pages (from-to)1-12
Number of pages12
JournalIn Vitro Cellular and Developmental Biology - Animal
DOIs
Publication statusAccepted/In press - 2016 Oct 3

Fingerprint

Zinc Fingers
Blastocyst
Stem cells
Zinc
Proteins
Genes
Embryonic Stem Cells
Mouse Embryonic Stem Cells
Genetic engineering
Pseudogenes
Genetic Engineering
Green Fluorescent Proteins
Transgenes
Mass spectrometry
Cluster Analysis
Mass Spectrometry
Nucleotides
Cell Line

Keywords

  • ES cell
  • Knock-in
  • Preimplantation embryo
  • Two-cell stage
  • Zscan4

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Expression analysis of the endogenous Zscan4 locus and its coding proteins in mouse ES cells and preimplantation embryos. / Ishiguro, Kei ichiro; Nakatake, Yuki; Chikazawa-Nohtomi, Nana; Kimura, Hiromi; Akiyama, Tomohiko; Oda, Mayumi; Ko, Shigeru; Ko, Minoru.

In: In Vitro Cellular and Developmental Biology - Animal, 03.10.2016, p. 1-12.

Research output: Contribution to journalArticle

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