Live imaging and tracking of genome regions in CRISPR/dCas9 knock-in mice

Genome Biol. 2018 Nov 8;19(1):192. doi: 10.1186/s13059-018-1530-1.

Abstract

CRISPR/dCas9 is a versatile tool that can be used to recruit various effectors and fluorescent molecules to defined genome regions where it can modulate genetic and epigenetic markers, or track the chromatin dynamics in live cells. In vivo applications of CRISPR/dCas9 in animals have been challenged by delivery issues. We generate and characterize a mouse strain with dCas9-EGFP ubiquitously expressed in various tissues. Studying telomere dynamics in these animals reveals surprising results different from those observed in cultured cell lines. The CRISPR/dCas9 knock-in mice provide an important and versatile tool to mechanistically study genome functions in live animals.

Keywords: CRISPR/dCas9; Genome labeling; Hepatocytes; Knock-in mice; Live imaging; Telomere dynamics; dCas9-EGFP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Cells, Cultured
  • Genome*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • HEK293 Cells
  • Hep G2 Cells
  • Hepatocytes / cytology
  • Hepatocytes / metabolism*
  • Humans
  • Image Processing, Computer-Assisted
  • Mice
  • Mice, Inbred C57BL
  • Molecular Imaging / methods*
  • Telomere / genetics
  • Telomere / metabolism*

Substances

  • Green Fluorescent Proteins