Rapid Disruption of Genes Specifically in Livers of Mice Using Multiplex CRISPR/Cas9 Editing

Gastroenterology. 2018 Dec;155(6):1967-1970.e6. doi: 10.1053/j.gastro.2018.08.037. Epub 2018 Aug 28.


Background & aims: Despite advances in gene editing technologies, generation of tissue-specific knockout mice is time-consuming. We used CRISPR/Cas9-mediated genome editing to disrupt genes in livers of adult mice in just a few months, which we refer to as somatic liver knockouts.

Methods: In this system, Fah-/- mice are given hydrodynamic tail vein injections of plasmids carrying CRISPR/Cas9 designed to excise exons in Hpd; the Hpd-edited hepatocytes have a survival advantage in these mice. Plasmids that target Hpd and a separate gene of interest can therefore be used to rapidly generate mice with liver-specific deletion of nearly any gene product.

Results: We used this system to create mice with liver-specific knockout of argininosuccinate lyase, which develop hyperammonemia, observed in humans with mutations in this gene. We also created mice with liver-specific knockout of ATP binding cassette subfamily B member 11, which encodes the bile salt export pump. We found that these mice have a biochemical phenotype similar to that of Abcb11-/- mice. We then used this system to knock out expression of 5 different enzymes involved in drug metabolism within the same mouse.

Conclusions: This approach might be used to develop new models of liver diseases and study liver functions of genes that are required during development.

Keywords: CRISPR/Cas9; Liver Gene Knockout; Mouse Models; SLiK.

Publication types

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

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 11 / genetics
  • Animals
  • Argininosuccinate Lyase / genetics*
  • CRISPR-Associated Protein 9 / administration & dosage*
  • CRISPR-Cas Systems / genetics*
  • Disease Models, Animal
  • Gene Editing / methods*
  • Hepatocytes / enzymology
  • Hepatocytes / physiology
  • Liver / enzymology*
  • Mice
  • Mice, Knockout
  • Oxidoreductases / genetics
  • Phenotype
  • Plasmids / genetics


  • ATP Binding Cassette Transporter, Subfamily B, Member 11
  • Hpd protein, mouse
  • Oxidoreductases
  • CRISPR-Associated Protein 9
  • Argininosuccinate Lyase