Methods and Compositions for Selecting siRNA of Improved Functionality

Abstract

Efficient sequence specific gene silencing is possible through the use of siRNA technology. By selecting particular siRNAs by rational design, one can maximize the generation of an effective gene silencing reagent, as well as methods for silencing genes. Methods, compositions, and kits generated through rational design of siRNAs are disclosed including those directed to nucleotide sequences for TTR.

1 . An isolated polynucleotide for use in gene silencing applications, wherein the polynucleotide is 18-30 bases and comprises at least 18 bases of SEQ ID NO:470 and comprises at least one nucleotide modification. 
     
     
         2 . An isolated polynucleotide for use in gene silencing applications, wherein the polynucleotide is 18-30 bases; and is complementary to SEQ ID NO:470; and comprises at least one nucleotide modification. 
     
     
         3 . A composition comprising the polynucleotide of  claim 1 . 
     
     
         4 . A composition comprising the polynucleotide of  claim 2 . 
     
     
         5 . An siRNA molecule comprising the polynucleotide of  claim 1 , wherein the siRNA molecule comprises a sense strand and an antisense strand. 
     
     
         6 . An siRNA molecule comprising the polynucleotide of  claim 2 , wherein the siRNA molecule comprises a sense strand and an antisense strand. 
     
     
         7 . The siRNA molecule of  claim 5 , wherein the siRNA molecule consists of: (a) a duplex region; and (b) either no overhang regions or at least one overhang region, wherein each overhang region contains six or fewer nucleotides, wherein the duplex region consists of a sense region and an antisense region, wherein the sense region and the antisense region together form the duplex region and the sense region and the antisense region are each 18-30 nucleotides in length and the antisense region comprises a sequence that is the complement of SEQ ID NO: 470. 
     
     
         8 . The siRNA molecule of  claim 7 , wherein the antisense region and the sense region are each 19-25 nucleotides in length. 
     
     
         9 . The siRNA molecule of  claim 8 , wherein the antisense region and the sense region are each 19 nucleotides in length. 
     
     
         10 . The siRNA molecule of  claim 7 , comprising no overhang region. 
     
     
         11 . The siRNA molecule of  claim 7 , comprising at least one overhang region. 
     
     
         12 . The antisense strand of the siRNA molecule of  claim 5 , comprising at least one nucleotide modification. 
     
     
         13 . The sense strand of the siRNA molecule of  claim 5 , comprising at least one nucleotide modification. 
     
     
         14 . The siRNA molecule of  claim 6 , wherein the siRNA molecule consists of: (a) a duplex region; and (b) either no overhang regions or at least one overhang region, wherein each overhang region contains six or fewer nucleotides, wherein the duplex region consists of a sense region and an antisense region, wherein the sense region and the antisense region together form the duplex region and the sense region and the antisense region are each 18-30 nucleotides in length and the antisense region comprises a sequence that is the complement of SEQ ID NO: 470. 
     
     
         15 . The siRNA molecule of  claim 14 , wherein the antisense region and the sense region are each 19-25 nucleotides in length. 
     
     
         16 . The siRNA molecule of  claim 15 , wherein the antisense region and the sense region are each 19 nucleotides in length. 
     
     
         17 . The siRNA molecule of  claim 14 , comprising no overhang region. 
     
     
         18 . The siRNA molecule of  claim 14 , comprising at least one overhang region. 
     
     
         19 . The antisense strand of the siRNA molecule of  claim 6 , comprising at least one nucleotide modification. 
     
     
         20 . The sense strand of the siRNA molecule of  claim 6 , comprising at least one nucleotide modification.