Original Article
Target gene knockout byCRISPR system in a mouse model of liver disease
Yue Ma, Li Mo, Jie Fang, Huanhuan Zhang, Chenhuan Yu
Published 2017-06-25
Cite as Inter J Epidemionl Infect Dis, 2017, 44(3): 151-155. DOI: 10.3760/cma.j.issn.1673-4149.2017.03.002
Abstract
ObjectiveTo construct a mouse model with liver-related disease by adeno-associated virus (AAV) -mediated gene transfer technology and CRISPR system.
MethodsAAV-green fluorescent protein (GFP) system (AAV-GFP) and CRISPR targeted gene knockout plasmid system (px330-sgGFP-Cas9) were constructed. Three groups of FVB/NJ mice were injected physiological saline (control group) , AAV-GFP (AAV-GFP group) and AAV-GFP+px330-sgGFP-Cas9 (sgGFP group) , respectively, by mouse tail vein hydrodynamic injection. Specific expression of GFP in the livers of mouse models were observed by Clairvivo OPT plus.
ResultsThe control group did not express GFP. AAV-GFP group began to express GFP after 72 h, and achieved a stable expression after 2 weeks. The sgGFP group expressed GFP on d9 and d12, and showed a significantly weakened expression after 2 weeks.
ConclusionsThe AAV-GFP system can stably express GFP in the liver of mice. CRISPR plasmid system can be used to knock down the expression of specific genes in mouse liver and establish a useful model of liver diseases.
Key words:
Plasmids; CRISPR; Adeno-associated virus system; Hydrodynamic injection
Contributor Information
Yue Ma
Zhejiang Key Laboratory of Experimental Animal and Safety Evaluation, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China
Li Mo
College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou 310053, China
Jie Fang
Zhejiang Key Laboratory of Experimental Animal and Safety Evaluation, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China
Huanhuan Zhang
Zhejiang Key Laboratory of Experimental Animal and Safety Evaluation, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China
Chenhuan Yu
Zhejiang Key Laboratory of Experimental Animal and Safety Evaluation, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China
