Modulation of mlck-l expression and uses thereof

Abstract

In various aspects and embodiments the invention provides methods and reagents for controlling gene expression, and for treating disorders and diseases. Embodiments provide methods and reagents specifically for the regulation of MLCK expression and for the use thereof in treating disorders and diseases. Various embodiments provide methods and reagents for specifically down regulating the expression of MLCK-L more efficiently than that of MLCK-S, and for the use thereof in treating disorders and diseases. Embodiments provide siNA for the same, particularly siRNAs. Various of the embodiments are useful for the treatment of inflammatory disorders and diseases, including, for one example in the regard, Asthma.

1 . A double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a myosin light chain kinase (MLCK) RNA, wherein: (a) each strand of said siNA molecule is about 19 to about 25 nucleotides in length; and (b) one strand of said siNA molecule comprises a region having a nucleotide sequence having sufficient complementarity to the MLCK RNA for the siNA molecule to direct cleavage of the MLCK RNA via RNA interference. 
     
     
         2 . A double-stranded siNA, according to  claim 1 , wherein the siNA is a siRNA. 
     
     
         3 . A double-stranded siNA according to  claim 1 , wherein the siNA cleaves MLCK-L RNA more efficiently than MLCK-S RNA. 
     
     
         4 . A double-stranded siNA, according to  claim 3 , wherein the siNA is a siRNA. 
     
     
         5 . A double-stranded siNA according to  claim 1 , wherein the siNA cleaves MLCK L RNA at least twice as efficiently as it cleaves MLCK-S RNA. 
     
     
         6 . A double-stranded siNA, according to  claim 5 , wherein the siNA is a siRNA. 
     
     
         7 . A double-stranded siRNA, according to  claim 1 , wherein the siNA specifically cleaves MLCK-L RNA and substantially does not cleave MLCK-S RNA. 
     
     
         8 . A double-stranded siNA, according to  claim 7 , wherein the siNA is a siRNA. 
     
     
         9 . A double-stranded siRNA, according to  claim 7 , wherein the siRNA comprises a region having the sequence of SEQ ID NO:3. 
     
     
         10 . A method for modulating MLCK expression in a cell, comprising introducing into a cell a double stranded siNA that cleaves MLCK RNA, wherein (a) each strand of said siNA molecule is about 19 to about 25 nucleotides in length; and (b) one strand of said siNA molecule comprises a region having a nucleotide sequence having sufficient complementarity to the MLCK RNA for the siNA molecule to direct cleavage of the MLCK RNA via RNA interference. 
     
     
         11 . A method according to  claim 10 , wherein the siNA specifically cleaves MLCK-L RNA and substantially does not cleave MLCK-S RNA. 
     
     
         12 . A method according to  claim 11 , wherein the siNA is a siRNA. 
     
     
         13 . A method according to  claim 12 , wherein the siRNA comprises a region having the sequence of SEQ ID NO:3. 
     
     
         14 . A method for modulating tissue inflammation in a patient suffering therefrom, comprising: administering to a patient suffering from tissue inflammation a nucleic acid in an amount and by a route effective to modulate said tissue inflammation, wherein said nucleic acid comprises a region complementary to an RNA encoded by an MLCK gene and is effective to direct cleavage specifically of said RNA by RNA interference. 
     
     
         15 . A method according to  claim 14 , wherein said nucleic acid is a double stranded siNA wherein (a) each strand of said siNA molecule is about 19 to about 25 nucleotides in length; and (b) one strand of said siNA molecule comprises a region having a nucleotide sequence having sufficient complementarity to the MLCK RNA for the siNA molecule to direct cleavage of the MLCK RNA via RNA interference. 
     
     
         16 . A method according to  claim 15 , wherein the siNA specifically cleaves MLCK-L RNA and substantially does not cleave MLCK-S RNA. 
     
     
         17 . A method according to  claim 16 , wherein the siNA is a siRNA. 
     
     
         18 . A method according to  claim 17 , wherein the siRNA comprises a region having the sequence of SEQ ID NO:3. 
     
     
         19 . A method for treating asthma in a patient suffering therefrom, comprising: administering to a patient suffering from asthma a nucleic acid in an amount and by a route effective to treat asthma, wherein said nucleic acid comprises a region complementary to an RNA encoded by an MLCK gene and is effective to direct cleavage specifically of said RNA by RNA interference. 
     
     
         20 . A method according to  claim 19 , wherein the nucleic acid is a siNA. wherein (a) each strand of said siNA molecule is about 19 to about 25 nucleotides in length; and (b) one strand of said siNA molecule comprises a region having a nucleotide sequence having sufficient complementarity to the MLCK RNA for the siNA molecule to direct cleavage of the MLCK RNA via RNA interference. 
     
     
         21 . A method according to  claim 20 , wherein the siNA specifically cleaves MLCK-L RNA and substantially does not cleave MLCK-S RNA. 
     
     
         22 . A method according to  claim 21 , wherein the siNA is a siRNA. 
     
     
         23 . A method according to  claim 22 , wherein the siRNA comprises a region having the sequence of SEQ ID NO:3.