Oligonucleotide Compositions with Enhanced Efficiency

Abstract

The oligonucleotide compositions of the present invention make use of combinations of oligonucleotides. In one aspect, the invention features an oligonucleotide composition including at least 2 different oligonucleotides targeted to a target gene. This invention also provides methods of inhibiting protein synthesis in a cell and methods of identifying oligonucleotide compositions that inhibit synthesis of a protein in a cell.

1 . An oligonucleotide composition comprising at least 3 different oligonucleotides targeted to at least three different nucleotide sequences within a target gene, wherein (i) the oligonucleotides bind to their target nucleotide sequence with high affinity and (ii) the oligonucleotides are GC enriched, and wherein the oligonucleotides are double-stranded RNA oligonucleotides. 
     
     
         2 . (canceled) 
     
     
         3 . (canceled) 
     
     
         4 . The oligonucleotide composition of  claim 1 , wherein the oligonucleotide compositions bind to their target nucleotide sequence with a Tm of at least about 60° C. 
     
     
         5 . The oligonucleotide composition of  claim 1 , wherein the oligonucleotides have a GC content of at least about 20%. 
     
     
         6 . The oligonucleotide composition of  claim 1 , wherein the composition comprises at least about 4 antisense oligonucleotides targeting at least four different nucleic acid sequences. 
     
     
         7 . The oligonucleotide composition of  claim 1 , wherein the composition comprises at least about 5 oligonucleotides targeting at least five different nucleic acid sequences. 
     
     
         8 . The oligonucleotide composition of  claim 1 , wherein the composition comprises at least about 6 oligonucleotides targeting at least six different nucleic acid sequences. 
     
     
         9 . The oligonucleotide composition of  claim 1 , wherein the oligonucleotides are at least about 25 nucleomonomers in length. 
     
     
         10 . The oligonucleotide composition of  claim 1 , wherein the oligonucleotides are greater than 25 nucleomonomers in length. 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . The oligonucleotide composition of  claim 1 , wherein at least one of the oligonucleotides comprise at least one modified internucleoside linkage. 
     
     
         14 . The oligonucleotide composition of  claim 1 , wherein at least one of the oligonucleotides comprise at least one modified sugar moiety. 
     
     
         15 . The oligonucleotide composition of  claim 1 , further comprising a pharmaceutically acceptable carrier. 
     
     
         16 . The oligonucleotide composition of  claim 1 , wherein the oligonucleotide composition achieves a level of inhibition of protein synthesis the same as or higher than the level of inhibition achieved by the most effective individual oligonucleotide of the composition. 
     
     
         17 . The oligonucleotide composition of  claim 1 , wherein the individual oligonucleotides are not separately tested for their ability to inhibit protein synthesis prior to their incorporation into the composition. 
     
     
         18 . The oligonucleotide composition of  claim 1 , wherein the oligonucleotide composition results in greater than about 80% inhibition of protein synthesis. 
     
     
         19 . A method of inhibiting protein synthesis in a cell comprising contacting the cell with at least 3 different oligonucleotides targeted to at least three different nucleotide sequences within a target gene, wherein (i) the oligonucleotides bind to their target nucleotide sequence with high affinity and (ii) the oligonucleotides are GC enriched, to thereby inhibit protein synthesis, and wherein the oligonucleotides are double-stranded RNA oligonucleotides. 
     
     
         20 . (canceled) 
     
     
         21 . (canceled) 
     
     
         22 . The method of  claim 19 , wherein the method is performed in a high-throughput format. 
     
     
         23 . A method of identifying function of a gene encoding a protein comprising: contacting the cell with at least 3 different oligonucleotides targeted to at least three different nucleotide sequences within a target gene, wherein (i) the oligonucleotides bind to their target nucleotide sequence with high affinity and (ii) the oligonucleotides are GC enriched, and assaying for a change in a detectable phenotype in the cell resulting from the inhibition of protein expression, to thereby determine the function of a gene, wherein the oligonucleotides are double-stranded RNA oligonucleotides. 
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . The method of  claim 23 , wherein the method is performed in a high-throughput format. 
     
     
         27 - 33 . (canceled)