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  • High-efficiency transduction of liver cancer cells by recombinant adeno-associated virus serotype 3 vectors.

High-efficiency transduction of liver cancer cells by recombinant adeno-associated virus serotype 3 vectors.

Journal of visualized experiments : JoVE (2011-03-30)
Chen Ling, Yuan Lu, Binbin Cheng, Katherine E McGoogan, Samantha W Y Gee, Wenqin Ma, Baozheng Li, George V Aslanidi, Arun Srivastava
ABSTRACT

Recombinant vectors based on a non-pathogenic human parvovirus, the adeno-associated virus 2 (AAV2) have been developed, and are currently in use in a number of gene therapy clinical trials. More recently, a number of additional AAV serotypes have also been isolated, which have been shown to exhibit selective tissue-tropism in various small and large animal models. Of the 10 most commonly used AAV serotypes, AAV3 is by far the least efficient in transducing cells and tissues in vitro as well as in vivo. However, in our recently published studies, we have documented that AAV3 vectors transduce human liver cancer - hepatoblastoma (HB) and hepatocellular carcinoma (HCC) - cell lines extremely efficiently because AAV3 utilizes human hepatocyte growth factor receptor as a cellular co-receptor for binding and entry in these cells. In this article, we describe the steps required to achieve high-efficiency transduction of human liver cancer cells by recombinant AAV3 vectors carrying a reporter gene. The use of recombinant AAV3 vectors carrying a therapeutic gene may eventually lead to the potential gene therapy of liver cancers in humans.

MATERIALS
Product Number
Brand
Product Description

Roche
Hexanucleotide Mix, solution, pkg of 100 μL, sufficient for 50 labeling reactions
Millipore
Benzonase® Nuclease, Purity > 99%, Effective viscosity reduction and removal of nucleic acids from protein solutions