In Vivo Editing of the Adult Mouse Liver Using CRISPR/Cas9 and Hydrodynamic Tail Vein Injection

Methods Mol Biol. 2019:1961:329-341. doi: 10.1007/978-1-4939-9170-9_20.

Abstract

CRISPR/Cas9 technology allows facile modification of the genome in virtually any desired way through the use of easily designed plasmid constructs that express a gRNA targeting a genomic site-of-interest and Cas9. Hydrodynamic tail vein injection, on the other hand, is a simple method to deliver "naked" plasmid DNA to 5-40% of the hepatocytes of the liver of adult mice. Here, we describe how these two techniques can be combined to create a workflow for fast, easy, and cost-efficient in vivo genome editing of the adult mouse liver. Using this method, large cohorts of mice with genetically modified livers can be established within 3 weeks to generate models for gene function in normal physiology and diseases of the liver.

Keywords: CRISPR/Cas9; Genetic mouse models; Hydrodynamic tail vein injection; Indels; Liver.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • Gene Editing*
  • Hepatocytes / metabolism
  • Liver / metabolism
  • Mice
  • Plasmids / genetics
  • RNA, Guide, CRISPR-Cas Systems / genetics

Substances

  • RNA, Guide, CRISPR-Cas Systems