CRISPR-Mediated Activation of Transposable Elements in Embryonic Stem Cells

Akihiko Sakashita, Masaru Ariura, Satoshi H. Namekawa

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages171-194
Number of pages24
DOIs
Publication statusPublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2509
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • CRISPR activation
  • Embryonic stem cells
  • Endogenous retroviruses
  • Germ cells
  • Meiosis
  • Spermatogenesis
  • Transposable elements

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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