Using piggyBac transposon gene expression vectors to transfect Zscan5b gene into mouse pluripotent stem cells

Mitsutoshi Yamada; Tohru Sugawara; Shinju Usami; Rina Nakanishi; Hidenori Akustu

doi:10.1016/j.xpro.2021.100811

Using piggyBac transposon gene expression vectors to transfect Zscan5b gene into mouse pluripotent stem cells

Mitsutoshi Yamada, Tohru Sugawara, Shinju Usami, Rina Nakanishi, Hidenori Akustu

Department of Obstetrics and Gynecology (Obstetrics)

Research output: Contribution to journal › Article › peer-review

Abstract

We recently demonstrated that the zygotic gene Zscan5b is involved in maintaining the genomic integrity of pluripotent stem cells (PSCs) during mitosis and that Zscan5b gene expression can prevent chromosomal abnormalities. Here, we provide a detailed protocol for the transfection of mouse embryonic stem cells (mESCs) and mouse-induced PSCs (miPSCs) with a Zscan5b constitutive expression plasmid, using the piggyBac transposon gene expression vector system. For complete details on the use and execution of this protocol, please refer to Ogawa et al. (2019).

Original language	English
Article number	100811
Journal	STAR Protocols
Volume	2
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2021.100811
Publication status	Published - 2021 Sep 17

Keywords

Cell Biology
Cell culture
Flow Cytometry/Mass Cytometry
Genomics
Molecular Biology
Single Cell
Stem Cells

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Neuroscience(all)
Immunology and Microbiology(all)
Medicine(all)

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10.1016/j.xpro.2021.100811

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title = "Using piggyBac transposon gene expression vectors to transfect Zscan5b gene into mouse pluripotent stem cells",

abstract = "We recently demonstrated that the zygotic gene Zscan5b is involved in maintaining the genomic integrity of pluripotent stem cells (PSCs) during mitosis and that Zscan5b gene expression can prevent chromosomal abnormalities. Here, we provide a detailed protocol for the transfection of mouse embryonic stem cells (mESCs) and mouse-induced PSCs (miPSCs) with a Zscan5b constitutive expression plasmid, using the piggyBac transposon gene expression vector system. For complete details on the use and execution of this protocol, please refer to Ogawa et al. (2019).",

keywords = "Cell Biology, Cell culture, Flow Cytometry/Mass Cytometry, Genomics, Molecular Biology, Single Cell, Stem Cells",

author = "Mitsutoshi Yamada and Tohru Sugawara and Shinju Usami and Rina Nakanishi and Hidenori Akustu",

note = "Funding Information: We thank A. Nakamura and T. Kawasaki for their technical support and laboratory management. M.Y. was supported in part by a KAKENHI grant from MEXT /JSPS ( JP20H03827 Kiban-B-20H03827 ) and a grant from the Takeda Science Foundation . Publisher Copyright: {\textcopyright} 2021 The Author(s)",

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doi = "10.1016/j.xpro.2021.100811",

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AU - Yamada, Mitsutoshi

AU - Sugawara, Tohru

AU - Usami, Shinju

AU - Nakanishi, Rina

AU - Akustu, Hidenori

N1 - Funding Information: We thank A. Nakamura and T. Kawasaki for their technical support and laboratory management. M.Y. was supported in part by a KAKENHI grant from MEXT /JSPS ( JP20H03827 Kiban-B-20H03827 ) and a grant from the Takeda Science Foundation . Publisher Copyright: © 2021 The Author(s)

PY - 2021/9/17

Y1 - 2021/9/17

N2 - We recently demonstrated that the zygotic gene Zscan5b is involved in maintaining the genomic integrity of pluripotent stem cells (PSCs) during mitosis and that Zscan5b gene expression can prevent chromosomal abnormalities. Here, we provide a detailed protocol for the transfection of mouse embryonic stem cells (mESCs) and mouse-induced PSCs (miPSCs) with a Zscan5b constitutive expression plasmid, using the piggyBac transposon gene expression vector system. For complete details on the use and execution of this protocol, please refer to Ogawa et al. (2019).

AB - We recently demonstrated that the zygotic gene Zscan5b is involved in maintaining the genomic integrity of pluripotent stem cells (PSCs) during mitosis and that Zscan5b gene expression can prevent chromosomal abnormalities. Here, we provide a detailed protocol for the transfection of mouse embryonic stem cells (mESCs) and mouse-induced PSCs (miPSCs) with a Zscan5b constitutive expression plasmid, using the piggyBac transposon gene expression vector system. For complete details on the use and execution of this protocol, please refer to Ogawa et al. (2019).

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KW - Cell culture

KW - Flow Cytometry/Mass Cytometry

KW - Genomics

KW - Molecular Biology

KW - Single Cell

KW - Stem Cells

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Using piggyBac transposon gene expression vectors to transfect Zscan5b gene into mouse pluripotent stem cells

Abstract

Keywords

ASJC Scopus subject areas

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