Simplified Purification of AAV and Delivery to the Pancreas by Intraductal Administration.

Simplified Purification of AAV and Delivery to the Pancreas by Intraductal Administration. Methods Mol Biol. 2019;1950:373-387 Authors: Guo P, Wiersch J, Xiao X, Gittes G Abstract 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. PMID: 30783986 [PubMed - indexed for MEDLINE]
Source: Mol Biol Cell - Category: Molecular Biology Authors: Tags: Methods Mol Biol Source Type: research