Cre重组酶
绿色荧光蛋白
转基因
胰腺
基因传递
Cre-Lox重组
生物
细胞生物学
基因
病毒载体
腺相关病毒
遗传增强
重组酶
分子生物学
载体(分子生物学)
计算生物学
转基因小鼠
遗传学
重组DNA
生物化学
重组
作者
Ping Guo,John Wiersch,Xiangwei Xiao,George K. Gittes
出处
期刊:Methods in molecular biology
日期:2019-01-01
卷期号:: 373-387
被引量:13
标识
DOI:10.1007/978-1-4939-9139-6_22
摘要
Genetic manipulation is a very powerful tool for studying diabetes, pancreatitis, and pancreatic cancer. Here we discuss the use of an adeno-associated virus (AAV) vector to modify gene expression, such as to introduce a green fluorescence protein (GFP) in wild-type mice, cre recombinase in loxP mice, or to inactivate a gene with shRNA. The use of viruses for genetic modification allows for time-specific genetic changes which have advantages over time-consuming and often complex cross-breeding strategies. Here we provide a detailed approach for this process from viral production and purification through pancreatic ductal infusion. Our protocol allows efficient delivery of AAV to mediate GFP or cre expression for cell lineage tracing in the mouse pancreas or for the delivery of transgenes under a specific promoter to these cells.
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