재조합 Adeno-associated Virus 제작과 효율적 신조적 감염

Abstract

Recombinant viral vectors based on the nonpathogenic parvovirus, adeno-associated virus (AAV), have a number of attractive features for the purpose of gene theraphy including the lack of cytotoxicity,

the ability to transduce non-dividing cells and its long-term transgene expression. In this study, we studied if recombinant AAV (rAAV-LacZ and rAAV-hIL10) vectors could efficiently transduce cells both in vivo and in vitro. We initially examined AAV-mediated gene transfer in HeLaRC32 cells in vitro. Expression of transgenes in rAAV was evaluated by RT-PCR (rAAV-hIL10) and X-gal staining. We then sudied if rAAV was effective in transducing renal tissue by direct injection of rAAV harboring the β–galactosidase gene (1×1010 physical particles/injected kidney) into the left kidney of mice. To assess gene transfer, the transduced renal tissue of mice was stained with X-gal. Based on visual assessment of X-gal staining, The expression of the β-galactosidase gene was mainly localized in tubular epithelial cells and Bowman’s capsular epithelial cells. The expression of the β-galactosidase gene in the renal tissue was detected for more than one month after the transduction of rAAV. These results suggest that rAAV vectors can be effective for transgene expression in the renal tissue.