Patent ID: 9181584
Filing Date: 2015-11-10
Classification: A61P,C12N,C12Q

Abstract:
1. A method for testing the expression and function of a small interfering RNA (siRNA) molecule, comprising: co-introducing into a mammalian cell (a) one or more vectors comprising (i) a first cassette comprising an RNA pol III promoter operatively linked to a first nucleic acid encoding a sense strand of a double stranded siRNA molecule and (ii) a second cassette comprising an RNA pol III promoter operatively linked to a second nucleic acid encoding an antisense strand of the double stranded siRNA molecule, wherein the first nucleic acid comprises a DNA sequence encoding the sense strand positioned at the first nucleotide of the transcript of the RNA pol III promoter and the second nucleic acid comprises a DNA sequence encoding the antisense strand positioned at the first nucleotide of the transcript of the RNA pol III promoter, wherein the first cassette and the second cassette are in a same vector or are in separate vectors and wherein the double stranded siRNA molecule is specific for a target sequence in a target gene, and (b) a vector comprising the target gene and a suitable second promoter; growing the mammalian cell under conditions suitable for expression of the nucleic acid sequences and the target gene; determining if the double stranded siRNA molecule is expressed and if expression of the target gene is reduced, whereby the expression and function of the siRNA molecule is tested.