Adenoviral delivered eGFP-intron splicing system for multiple gene RNAi

An eGFP-intron splicing system that allows for co-ordinated expression of up to four siRNAs from a single adenoviral vector has been developed. In this splicing structure the intron, embedded by a multiple miR30-based shRNAs, is located between two incomplete eGFP domains which require successful splicing for functionality. To prove the principle of the method, an adenoviral vector delivering four transcripts targeting survivin, XIAP, Hec1, and VEGF was developed which enabled the knockdown of target genes by 70, 70, 54 and 44%, respectively, in HeLa cells. This is the first report of multi-siRNA engineering technology in the context of adenoviral vector which would enable concomitant knockdown of tumor-related target genes. The results provide a strategy for gene function analysis and cancer gene therapy.