Gene Editing of Mouse Embryonic and Epiblast Stem Cells

Methods Mol Biol. 2019:1940:77-95. doi: 10.1007/978-1-4939-9086-3_6.

Abstract

Efficient and reliable methods for gene editing are critical for the generation of loss-of-gene function stem cells and genetically modified mice. Here, we outline the application of CRISPR-Cas9 technology for gene editing in mouse embryonic stem cells (mESCs) to generate knockout ESC chimeras for the fast-tracked analysis of gene function. Furthermore, we describe the application of gene editing directly to mouse epiblast stem cells (mEpiSCs) for modelling germ layer differentiation in vitro.

Keywords: CRISPR-Cas9; Embryonic stem cells; Epiblast stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Cells, Cultured
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Frameshift Mutation / genetics*
  • Gene Editing / methods*
  • Gene Knockout Techniques / methods*
  • Germ Layers / cytology*
  • Mice
  • Mouse Embryonic Stem Cells / cytology*
  • Plasmids / genetics
  • RNA, Guide, CRISPR-Cas Systems / genetics

Substances

  • RNA, Guide, CRISPR-Cas Systems