Unknown

Dataset Information

0

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


ABSTRACT: 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.

SUBMITTER: Pankowicz FP 

PROVIDER: S-EPMC6420307 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature

altmetric image

Publications


<h4>Background & aims</h4>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.<h4>Methods</h4>In this system, Fah<sup>-/-</sup> 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 adva  ...[more]

Similar Datasets

| S-EPMC6180501 | biostudies-literature
| S-EPMC5512712 | biostudies-literature
| S-EPMC9832834 | biostudies-literature
| S-EPMC8747517 | biostudies-literature
| S-EPMC6319321 | biostudies-literature
| S-EPMC5940154 | biostudies-literature
| S-EPMC4371917 | biostudies-literature
| S-EPMC5935052 | biostudies-literature
| S-EPMC6921047 | biostudies-literature
| S-EPMC4265475 | biostudies-literature