Patent ID: 8299045

Claim:
A method for expressing an interfering RNA molecule in a mammalian cell, comprising: (a) introducing into a mammalian cell an expression cassette comprising an adenoviral VA1 gene and a nucleic acid encoding an interfering RNA (RNAi) precursor molecule, wherein the adenoviral VA1 gene comprises the adenoviral VA1 promoter and a coding sequence for the VA1 RNA, wherein the nucleic acid is inserted at a BstEII site within a non-essential stem region of the adenoviral VA1 coding sequence, wherein the nucleic acid encoding the RNAi precursor molecule encodes a hairpin siRNA (shRNA) or a precursor microRNA (precursor miRNA); (b) maintaining the mammalian cell produced in step (a) under conditions and for a time sufficient to express the VA1 RNA containing the RNAi precursor molecule and to cleave the RNAi precursor molecule out of the VA1 RNA; and (c) maintaining the mammalian cell produced in step (b) under conditions and for a time sufficient for Dicer to cleave the RNAi precursor molecule to produce an interfering RNA molecule, wherein the RNA interfering molecule is an siRNA or a miRNA.