Targeting human transgenes

Adeno-associated virus vectors can be used for efficient transgene insertion into targeted loci in human cells.

Jonathan Weitzman(jonathanweitzman@hotmail.com)
Jul 1, 2002

Adeno-associated viruses (AAVs) are single-stranded DNA vectors that have shown promise as gene-targeting vectors for experimental and therapeutic applications. In the July issue of Nature Biotechnology, Roli Hirata and colleagues at the University of Washington, Seattle, describe a way of using AAV to introduce a functional transgene cassette into defined genomic loci in human cells in culture (Nat Biotechnol 2002, 20:735-738).

The AAV vectors can deliver gene cassettes of up to 1.5 kb. Hirata et al. designed an AAV vector containing a selectable neomycin cassette within the hypoxanthine phosphoribosyl transferase (HPRT) gene. They then infected diploid male human fibroblasts, or HT1080 fibrosarcoma cells, and selected for neomycin resistance and functional HPRT expression. The targeting efficiency was as high as 1% of the total cell population. Hirata et al. also used a similar strategy to disrupt the autosomal type I collagen (COL1A1)...

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