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

Gastroenterology
Francis P PankowiczKarl-Dimiter Bissig

Abstract

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. 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. 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. This approach might be used to develop new models of liver diseases and study liver functions of ...Continue Reading

References

Aug 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·P C OrbanJ D Marth
Jul 25, 1992·Proceedings of the National Academy of Sciences of the United States of America·M LaksoH Westphal
Feb 1, 2003·Molecular Genetics and Metabolism·V Reid SuttonWilliam J Craigen
Jan 14, 2012·Genetics in Medicine : Official Journal of the American College of Medical Genetics·Sandesh C S NagamaniBrendan Lee
Sep 30, 2015·Nature Communications·Yuanyuan ZhangJohn D Schuetz
Aug 31, 2016·Nature Communications·Francis P PankowiczKarl-Dimiter Bissig
Jul 1, 2017·Nature Communications·Mercedes BarziKarl-Dimiter Bissig

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Citations

May 31, 2019·World Journal of Gastroenterology : WJG·Viktor LimanskiyDinesh Vyas
Jan 21, 2021·International Journal of Molecular Sciences·Muhammad Imran SohailPeter Chiba
Nov 23, 2021·Applied Microbiology and Biotechnology·Yali KongWei-Min Zhang

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