Recombinant adeno-associated virus expressing human antisense gene CyP2J2 and its preparation methods

Abstract

A recombinant adeno-associated virus (rAAV) expressing antisense human CYP2J2 gene produced by a method of co-transfection into 293 cell by calcium phosphate precipitation, the rAAV having the following three plasmids: a) pXX2: a packing plasmid having a nucleotide sequence encoding AAV Rep and Cap protein, wherein the nucleotide sequence is operably linked upstream and downstream to a p5 promoter respectively to increase expression 15-fold; b) pXXUF1-anti2J2: an eukaryotic expression vector having a CMV promoter, a NotI site for insertion of target genes, the CYP2J2 gene oriented in the 3′ to 5′ direction, and inverted terminal repeats (ITRs); and c) pXX6: a helper plasmid deleted of the pathogenic gene sequences of adenovirus and having an E1A, an E2A, and a VA1 RNA gene of adenovirus. Also provided is a method of preparing the rAAV.

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