CRISPR-Mediated Activation of Transposable Elements in Embryonic Stem Cells

Methods Mol Biol. 2022:2509:171-194. doi: 10.1007/978-1-0716-2380-0_11.

Abstract

Mounting evidence has established that subsets of transposable elements (TEs) function as gene regulatory elements in a cell type- and species-specific manner. Here we describe an in vitro system to ectopically activate TEs using CRISPR-mediated activation (CRISPRa) for functional studies in mouse embryonic stem cells (ESCs). We established a stable mouse CRISPRa ESC line, in which expression of guide RNA enables the activation of TE-derived enhancers and the expression of their adjacent genes. We show an example of ectopic activation of TE-derived enhancers that function in male meiosis, as well as the expression of adjacent germline genes in ESCs. This system can also be applied to functional studies of TEs that are not active in ESCs.

Keywords: CRISPR activation; Embryonic stem cells; Endogenous retroviruses; Germ cells; Meiosis; Spermatogenesis; Transposable elements.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • DNA Transposable Elements* / genetics
  • Embryonic Stem Cells / metabolism
  • Male
  • Mice
  • Mouse Embryonic Stem Cells
  • Regulatory Sequences, Nucleic Acid

Substances

  • DNA Transposable Elements