Patent ID: 8097715

Claim:
A method for making a multitargeting interfering RNA molecule, comprising the steps of: a) selecting one or more target RNA molecules, wherein the modulation in expression of the target RNA molecules is desired; b) obtaining at least one nucleotide sequence for each of the target RNA molecules; c) selecting a length, n, in nucleotides, for a seed sequence, wherein n=about 6 or more; d) obtaining a collection of candidate seed sequences of the length n from each nucleotide sequence obtained in step b), wherein a candidate seed sequence and its complete complement are not palindromic, and the candidate seed sequence occurs at least once in one or more of the nucleotide sequences obtained in step b), and its complete complement occurs at least once in one or more of the nucleotide sequences obtained in step b); e) determining the genetic context of each of the candidate seed sequence and its complete complement, by collecting, for each occurrence of the candidate seed sequence and its complete complement, 5′ and 3′ flanking sequence of each of the candidate seed sequences; f) selecting a seed sequence of the length n from the collection of candidate seed sequences obtained in step d); g) selecting a first consensus target sequence, which comprises the seed sequence selected in step f) and a consensus 3′-flanking sequence to a seed sequence determined from the sequences obtained in step b); h) selecting a second consensus target sequence, which comprises the complete complement of the seed sequence and a consensus 3′-flanking sequence to the complete complement of the seed sequence determined from the sequences obtained in step b); i) obtaining a first strand sequence, which comprises the first consensus target sequence selected in step g) and, adjacent to and connected with the 5′-end of the first consensus target sequence, a complement of the consensus 3′ flanking sequence of step h); j) obtaining a second strand sequence which comprises the second consensus target sequence selected in step h) and, adjacent to and connected with the 5′-end of the second consensus target sequence, a complement of the consensus 3′ flanking sequence of step g), and; k) making a multitargeting interfering RNA molecule comprising a first strand having the first strand sequence in step i) and a second strand having the second strand sequence obtained in step j), wherein the multitargeting interfering RNA molecule targets VEGF-A.