US2006142228A1PendingUtilityA1

Methods and compositions concerning siRNA's as mediators of RNA interference

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Assignee: AMBION INCPriority: Dec 23, 2004Filed: Dec 23, 2004Published: Jun 29, 2006
Est. expiryDec 23, 2024(expired)· nominal 20-yr term from priority
Inventors:Lance FordJoseph Krebs
C12N 15/1137C12Y 502/01008C12Y 102/01012C12N 2330/30C12N 2310/14C12N 15/111
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Claims

Abstract

The present invention concerns an isolated siRNA of from about 5 to about 20 nucleotides that mediates RNA interference. Also disclosed are methods of reducing expression of a target gene in a cell comprising obtaining at least one siRNA of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 basepairs in length; and delivering the siRNA into the cell. The siRNAs can be chemically synthesized RNA or an analog of a naturally occurring RNA.

Claims

exact text as granted — not AI-modified
1 . An isolated RNA of from about 5 to about 20 nucleotides that mediates RNA interference of a target mRNA.  
     
     
         2 . The isolated RNA of  claim 1  further comprising a terminal 3′ hydroxyl group.  
     
     
         3 . The isolated RNA of  claim 1  which is chemically synthesized RNA or an analog of a naturally occurring RNA.  
     
     
         4 . The isolated RNA of  claim 1 , wherein the RNA is from about 12 to about 18 nucleotides in length.  
     
     
         5 . The isolated RNA of  claim 1 , wherein the RNA is from about 14 to about 16 nucleotides in length.  
     
     
         6 . The isolated RNA of  claim 1 , wherein the RNA is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides in length.  
     
     
         7 . The isolated RNA of  claim 1 , wherein the RNA is an siRNA.  
     
     
         8 . The isolated RNA of  claim 1 , wherein the RNA is a single-stranded.  
     
     
         9 . The isolated RNA of  claim 1 , wherein the RNA is a double-stranded.  
     
     
         10 . The isolated RNA of  claim 9 , wherein the 3′ ends of the double stranded RNA comprises a 2, 3, 4, or 5 nucleotide overhang.  
     
     
         11 . The isolated RNA of  claim 10 , wherein the nucleotide overhang is a 2 nucleotide overhang.  
     
     
         12 . The isolated RNA of  claim 11 , wherein the 2 nucleotides are thymine.  
     
     
         13 . The isolated RNA of  claim 9 , wherein the 5′ ends of the double stranded RNA comprises a 2, 3, 4, or 5 nucleotide overhang.  
     
     
         14 . The isolated RNA of  claim 13 , wherein the nucleotide overhang is a 2 nucleotide overhang.  
     
     
         15 . The isolated RNA of  claim 14 , wherein the 2 nucleotides are thymine.  
     
     
         16 . The isolated RNA of  claim 1 , wherein the RNA is formulated into a pharmaceutically acceptable composition.  
     
     
         17 . The isolated RNA of  claim 1 , wherein the RNA associates with a protein complex.  
     
     
         18 . The isolated RNA of  claim 17 , wherein the protein complex is RNA-induced silencing complex (RISC).  
     
     
         19 . The isolated RNA of  claim 1 , wherein the isolated RNA comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13; SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17; SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30; SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO 35, SEQ ID NO: 36; SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, and SEQ ID NO: 54.  
     
     
         20 . A method of reducing expression of a target gene in a cell comprising: 
 a) obtaining at least one siRNA of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides in length; and    b) delivering the siRNA into the cell.    
     
     
         21 . The method of  claim 20 , wherein the siRNA is 10, 11, 12, 13, 14, 15, 16, or 17 nucleotides in length.  
     
     
         22 . The method of  claim 21 , wherein the siRNA is 12, 13, 14, or 15 nucleotides in length.  
     
     
         23 . The method of  claim 20 , wherein the siRNA is chemically synthesized siRNA or an analog of a naturally occurring siRNA.  
     
     
         24 . The method of  claim 20 , further comprising isolating the siRNA prior to delivery.  
     
     
         25 . The method of  claim 20 , further comprising obtaining at least two siRNAs and delivering them into the cell.  
     
     
         26 . The method of  claim 20 , further comprising obtaining a pool of siRNAs and delivering the pool into the cell.  
     
     
         27 . The method of  claim 20 , wherein the cell is comprised in an organism.  
     
     
         28 . The method of  claim 20 , wherein the cell is a human cell.  
     
     
         29 . The method of  claim 20 , wherein multiple siRNA molecules are delivered into the cell.  
     
     
         30 . The method of  claim 20 , wherein the siRNA comprises a terminal 3′ hydroxyl group.  
     
     
         31 . The method of  claim 20 , wherein the siRNA is double-stranded.  
     
     
         32 . The method of  claim 31 , wherein the 3′ ends of the double stranded siRNA comprise a 2, 3, 4, or 5 nucleotide overhang.  
     
     
         33 . The method  claim 32 , wherein the nucleotide overhang is a 2 nucleotide overhang.  
     
     
         34 . The method of  claim 33 , wherein the 2 nucleotides are thymine.  
     
     
         35 . The method of  claim 31 , wherein the 5′ ends of the double stranded siRNA comprise a 2, 3, 4, or 5 nucleotide overhang.  
     
     
         36 . The method of  claim 35 , wherein the nucleotide overhang is a 2 nucleotide overhang.  
     
     
         37 . The method of  claim 36 , wherein the two nucleotides are thymine.  
     
     
         38 . The method of  claim 20 , wherein the siRNA is formulated into a pharmaceutically acceptable composition.  
     
     
         39 . The method of  claim 20 , wherein the siRNA associates with a protein complex.  
     
     
         40 . The method of  claim 39 , wherein the protein complex is RNA-induced silencing complex (RISC).  
     
     
         41 . The method of  claim 20 , wherein the siRNA comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13; SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17; SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30; SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO 35, SEQ ID NO: 36; SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, and SEQ ID NO: 54.

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