Rapid Disruption of Genes Specifically in Livers of Mice Using Multiplex CRISPR/Cas9 Editing
| Autor: | Pankowicz, Francis P, Barzi, Mercedes, Kim, Kang Ho, Legras, Xavier, Martins, Celeste Santos, Wooton-Kee, Clavia Ruth, Lagor, William R, Marini, Juan C, Elsea, Sarah H, Bissig-Choisat, Beatrice, Moore, David D, Bissig, Karl-Dimiter |
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| Rok vydání: | 2018 |
| Předmět: |
Liver Gene Knockout
Knockout ATP Binding Cassette Transporter Chronic Liver Disease and Cirrhosis Clinical Sciences Mouse Models Paediatrics and Reproductive Medicine Mice Rare Diseases CRISPR-Associated Protein 9 Genetics Animals SLiK CRISPR/Cas9 Gene Editing Gastroenterology & Hepatology Animal Liver Disease Neurosciences Argininosuccinate Lyase Phenotype Liver Subfamily B Disease Models Hepatocytes Member 11 CRISPR-Cas Systems Oxidoreductases Digestive Diseases Plasmids Biotechnology |
| Zdroj: | Gastroenterology, vol 155, iss 6 |
| Popis: | Background & aimsDespite 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.MethodsIn 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.ResultsWe 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.ConclusionsThis approach might be used to develop new models of liver diseases and study liver functions of genes that are required during development. |
| Databáze: | OpenAIRE |
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