Rapid Disruption of Genes Specifically in Livers of Mice Using Multiplex CRISPR/Cas9 Editing.
Published
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
Digital Object Identifier (DOI)
- 10.1053/j.gastro.2018.08.037
Language
- eng
Conference Location
- United States