Patent ID: 7422896

Claim:
A DNA-mediated silencing of gene (DMSG) cassette, comprising, in operative linkage, a distal sequence element (DSE), an RNA polymerase III (pol III) promoter, an expressible template nucleotide sequence, and at least one pol III terminator, wherein the terminator comprises SEQ ID NO:50, wherein the DSE is about −79 to −72 nucleotides upstream of the expressible template nucleotide sequence, wherein the expressible template nucleotide sequence is heterologous with respect to the pol III promoter, wherein the expressible template nucleotide sequence consists of at least about 16 nucleotides encoding an intermediate small interfering ribonucleic acid (RNA) molecule (siRNA), which mediates RNA interference of a target RNA, said intermediate siRNA comprising: a) a 5′ portion, which comprises at least about 15 nucleotides complementary to a sense strand of the target RNA, and, optionally, a 3′ terminal portion, comprising about 1 to 5 nucleotides, wherein the intermediate siRNA selectively hybridizes to the sense strand of the target RNA; or b) a 5′ portion, which comprises at least about 15 nucleotides complementary to an antisense strand of the target RNA, and, optionally, a 3′ terminal portion, comprising about 1 to 5 nucleotides, wherein the intermediate siRNA selectively hybridizes to the antisense strand of the target RNA, and wherein the encoded intermediate siRNA does not activate PKR in a mammalian cell.