Method and composition for reducing the expression of rock-ii

Abstract

Methods and compositions for reducing the expression of Rho-associated, coiled-coil containing protein kinase 2 (ROCK-II) are provided.

1 . A nucleic acid molecule that directs the cleavage of a Rho-associated, coiled-coil containing protein kinase 2 (ROCK-II)-encoding RNA transcript via RNA interference (RNAi), comprising a first nucleotide sequence and a second nucleotide sequence, wherein:
 said first nucleotide sequence comprises SEQ ID NO:11 or SEQ ID NO:29;   said second nucleotide sequence is substantially complementary to said first nucleotide sequence;   wherein each of said first and second nucleotide sequences is about 21 nucleotides in length, and sufficiently complementary to the other nucleotide sequence to allow said first and second nucleotide sequences to anneal and form a duplex comprising at least 19 basepairs; and   wherein said second nucleotide sequence is sufficiently complementary to SEQ ID NO:2 for the nucleic acid molecule to direct cleavage of said ROCK-II-encoding RNA transcript and decrease the expression of ROCK-II via RNAi.   
     
     
         2 . The nucleic acid of  claim 1 , wherein:
 said first nucleotide sequence occurs within a first strand of RNA;   said second nucleotide sequence occurs within a second strand of RNA;   each of said first and second nucleotide sequences is about 21 nucleotides in length; and   said first strand of RNA and said second strand of RNA are annealed to form an intermolecular duplex.   
     
     
         3 . A composition comprising the nucleic acid of  claim 2  and a pharmaceutically acceptable carrier or diluent. 
     
     
         4 . The composition of  claim 3 , further comprising an uptake agent. 
     
     
         5 . The composition of  claim 4 , wherein said uptake agent is selected from Lipofectin®, Lipofectamine™, and Cellfectin®. 
     
     
         6 . A modified equivalent of the nucleic acid of  claim 2 . 
     
     
         7 . A composition comprising the modified equivalent of  claim 6  and a pharmaceutically acceptable carrier or diluent. 
     
     
         8 . The composition of  claim 7 , further comprising an uptake agent. 
     
     
         9 . The composition of  claim 8 , wherein said uptake agent is selected from Lipofectin®, Lipofectamine™, and Cellfectin®. 
     
     
         10 . The nucleic acid of  claim 1 , wherein
 said first and second nucleotide sequences each further comprise two 2′-deoxythymidine residues at their 3′ ends, and   said nucleic acid is prepared by the annealing of the nucleic acids of SEQ ID NOs:54 and 55, or SEQ ID NOs:68 and 69.   
     
     
         11 . The nucleic acid of  claim 1 , wherein:
 said first nucleotide sequence and said second nucleotide sequence occur within a single strand of RNA,   said single strand of RNA is comprised of about 52 nucleotides,   and said first nucleotide sequence and said second nucleotide sequence anneal to form an intramolecular duplex of about 19 basepairs.   
     
     
         12 . A composition comprising the nucleic acid of  claim 11  and a pharmaceutically acceptable carrier or diluent. 
     
     
         13 . The composition of  claim 12 , further comprising an uptake agent. 
     
     
         14 . The composition of  claim 13 , wherein said uptake agent is selected from Lipofectin®, Lipofectamine™, and Cellfectin®. 
     
     
         15 . The nucleic acid of  claim 11 , wherein said single strand of RNA comprises SEQ ID NO:76 or SEQ ID NO: 83. 
     
     
         16 . A composition comprising the nucleic acid of  claim 15  and a pharmaceutically acceptable carrier or diluent. 
     
     
         17 . The composition of  claim 16 , further comprising an uptake agent. 
     
     
         18 . The composition of  claim 17 , wherein said uptake agent is selected from Lipofectin®, Lipofectamine™, and Cellfectin®. 
     
     
         19 . A method of reducing ROCK-II expression in a human cell comprising contacting the cell with an effective amount of the composition of  claim 4 . 
     
     
         20 . A method of reducing ROCK-II expression in a human cell comprising contacting the cell with an effective amount of the composition of  claim 8 . 
     
     
         21 . A method of reducing ROCK-II expression in a human cell comprising contacting the cell with an effective amount of the composition of  claim 13 . 
     
     
         22 . A method of reducing ROCK-II expression in a human cell comprising contacting the cell with an effective amount of the composition of  claim 17 .