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

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Pankowicz, FP; Barzi, M; Kim, KH; Legras, X; Martins, CS; Wooton-Kee, CR; Lagor, WR; Marini, JC; Elsea, SH; Bissig-Choisat, B; Moore, DD; Bissig, K-D

Published Date

  • December 2018

Published In

Volume / Issue

  • 155 / 6

Start / End Page

  • 1967 - 1970.e6

PubMed ID

  • 30170115

Pubmed Central ID

  • 30170115

Electronic International Standard Serial Number (EISSN)

  • 1528-0012

International Standard Serial Number (ISSN)

  • 0016-5085

Digital Object Identifier (DOI)

  • 10.1053/j.gastro.2018.08.037

Language

  • eng