Functional and hyperfunctional siRNA

Abstract

Efficient sequence specific gene silencing is possible through the use of siRNA technology. By selecting particular siRNAs by rationale design, one can maximize the generation of an effective gene silencing reagent, as well as methods for silencing genes.

1 .- 19 . (canceled)  
     
     
         20 . A kit comprised of a first siRNA and a second siRNA, wherein said first siRNA comprises 18-30 base pairs that form a first sense strand and a first antisense strand, wherein said first antisense strand is at least substantially complementary to a first region of a target, said second siRNA comprises 18-30 base pairs that form a second sense strand and a second antisense strand, wherein said second antisense strand is at least substantially complementary to a second region of said target, said first siRNA has an internal stability at said first siRNA's 3′ sense end that is lower than the internal stability of said first siRNA's 5′ sense end, and said second siRNA has an internal stability at second siRNA's 3′ sense end that is lower than the internal stability of said second siRNA's 5′ sense end.  
     
     
         21 . The kit of claim  1 , further comprising a third siRNA, wherein said third siRNA comprises 18-30 base pairs that form a third sense strand and a third antisense strand, wherein said third antisense strand is at least substantially complementary to a third region of said target, and said third siRNA has an internal stability at said third siRNA's 3′ sense end that is lower than the internal stability of said third siRNA's 5′ sense end.  
     
     
         22 . The kit of claim  2 , further comprising a fourth siRNA, wherein said fourth siRNA comprises 18-30 base pairs that form a fourth sense strand and a fourth antisense strand, wherein said fourth antisense strand is at least substantially complementary to a fourth region of said target, and said fourth siRNA has an internal stability at said fourth siRNA's 3′ sense end that is lower than the internal stability of said fourth siRNA's 5′ sense end.  
     
     
         23 . The kit of claim  1 , wherein the base at position 19 of the first sense strand is A.  
     
     
         24 . The kit of claim  4 , wherein the base at position 19 of the second sense strand is A.  
     
     
         25 . The kit of claim  2 , wherein the base at position 19 of the first sense strand is A, the base at position 19 of the second sense strand is A, and the base at position 19 of the third sense strand is A.  
     
     
         26 . The kit of claim  3 , wherein the base at position 19 of the first sense strand is A, the base at position 19 of the second sense strand is A, the base at position 19 of the third sense strand is A and the base at position 19 of the fourth sense strand is A.  
     
     
         27 . The kit of claim  1 , wherein the base at position 1 of the first sense strand is G.  
     
     
         28 . The kit of claim  8 , wherein the base at position 1 of the second sense strand is G.  
     
     
         29 . The kit of claim  2 , wherein the base at position 1 of the first sense strand is G, the base at position 1 of the second sense strand is G, and the base at position 1 of the third sense strand is G.  
     
     
         30 . The kit of claim  3 , wherein the base at position 1 of the first sense strand is G, the base at position I of the second sense strand is G, the base at position 1 of the third sense strand is G and the base at position 1 of the fourth sense strand is G.  
     
     
         31 . The kit of claim  4 , wherein the base at position 1 of the first sense strand is G.  
     
     
         32 . The kit of claim  5 , wherein the base at position 1 of the first sense strand and second sense strand is G.  
     
     
         33 . The kit of claim  6 , wherein the base at position 1 of the first sense strand is G, the base at position 1 of the second sense strand is G, and the base at position 1 of the third sense strand is G.  
     
     
         34 . The kit of claim  7 , wherein the base at position 1 of the first sense strand is G, the base at position 1 of the second sense strand is G, the base at position 1 of the third sense strand is G and the base at position I of the fourth sense strand is G.  
     
     
         35 . A kit comprised of a first siRNA and a second siRNA, wherein said first siRNA comprises 18-30 base pairs that form a first sense strand and a first antisense strand, wherein said first antisense strand is at least substantially complementary to a first region of a target, said second siRNA comprises 18-30 base pairs that form a second sense strand and a second antisense strand, wherein said second antisense strand is at least substantially complementary to a second region of said target, said first siRNA has an internal stability at said first siRNA's 3′ sense end that is lower than the internal stability of said first siRNA's 5′ sense end, said second siRNA has an internal stability at second siRNA's 3′ sense end that is lower than the internal stability of said second siRNA's 5′ sense end, and said first sense siRNA and said second sense siRNA each satisfy at least one of the sequence criteria selected from the group consisting of: G is the base at position 1, G is the base at position 2, A is the base at position 3, A is the base at position 6, C is the base at position 9, U is the base at position 10, A is the base at position 13, and A is the base at position 19.  
     
     
         36 . The kit of claim  16 , wherein said first sense siRNA and said second sense siRNA each satisfy at least two of said criteria.  
     
     
         37 . The kit of claim  17 , wherein said first sense siRNA and said second sense siRNA each satisfy at least three of said criteria.  
     
     
         38 . The kit of claim  18 , wherein said first sense siRNA and said second sense siRNA each satisfy at least four of said criteria.  
     
     
         39 . The kit of claim  19 , wherein said first sense siRNA and said second sense siRNA each satisfy at least five of said criteria.  
     
     
         40 . The kit of  claim 20 , wherein said first sense siRNA and said second sense siRNA each satisfy at least six of said criteria.  
     
     
         41 . The kit of  claim 21 , wherein said first sense siRNA and said second sense siRNA each satisfy at least seven of said criteria.  
     
     
         42 . The kit of  claim 22 , wherein said first sense siRNA and said second sense siRNA each satisfy at least eight of said criteria.  
     
     
         43 . The kit of claim  16 , further comprising a third siRNA, wherein said third siRNA comprises 18-30 base pairs that form a third sense strand and a third antisense strand, wherein said third antisense strand is at least substantially complementary to a third region of said target, and said third siRNA has an internal stability at said third siRNA's 3′ sense end that is lower than the internal stability of said siRNA's 5′ sense end, wherein said third sense strand satisfies at least one of said criteria.  
     
     
         44 . The kit of claim  17 , wherein said third sense strand satisfies at least two of said criteria.  
     
     
         45 . The kit of claim  18 , wherein said third sense strand satisfies at least three of said criteria.  
     
     
         46 . The kit of claim  19 , wherein said third sense strand satisfies at least four of said criteria.  
     
     
         47 . The kit of  claim 20 , wherein said third sense strand satisfies at least five of said criteria.  
     
     
         48 . The kit of  claim 21 , wherein said third sense strand satisfies at least six of said criteria.  
     
     
         49 . The kit of  claim 22 , wherein said third sense strand satisfies at least seven of said criteria.  
     
     
         50 . The kit of  claim 23 , wherein said third sense strand satisfies at least seven of said criteria.  
     
     
         51 . The kit of  claim 24 , further comprising a fourth siRNA, wherein said fourth siRNA comprises 18-30 bases pairs that form a fourth sense strand and a fourth antisense strand, wherein said fourth antisense strand is at least substantially complementary to a fourth region said target, and said fourth siRNA has an internal stability at said fourth siRNA's 3′ sense end that is lower than the internal stability of said siRNA's 5′ sense end, wherein said fourth siRNA satisfies at least one of said criteria.  
     
     
         52 . The kit of  claim 31 , wherein said third sense strand satisfies at least two of said criteria.  
     
     
         53 . The kit of  claim 32 , wherein said third sense strand satisfies at least three of said criteria.  
     
     
         54 . The kit of  claim 33 , wherein said third sense strand satisfies at least four of said criteria.  
     
     
         55 . The kit of  claim 34 , wherein said third sense strand satisfies at least five of said criteria.  
     
     
         56 . The kit of  claim 35 , wherein said third sense strand satisfies at least six of said criteria.  
     
     
         57 . The kit of  claim 36 , wherein said third sense strand satisfies at least seven of said crit