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

STAR Protoc. 2021 Sep 11;2(3):100811. doi: 10.1016/j.xpro.2021.100811. eCollection 2021 Sep 17.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA-Binding Proteins / genetics
  • Female
  • Gene Expression / genetics
  • Genetic Vectors / genetics*
  • Induced Pluripotent Stem Cells* / cytology
  • Induced Pluripotent Stem Cells* / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mouse Embryonic Stem Cells* / cytology
  • Mouse Embryonic Stem Cells* / metabolism
  • Plasmids / genetics*
  • Transfection / methods*
  • Zinc Fingers / genetics

Substances

  • DNA-Binding Proteins