Human RNase H and compositions and uses thereof

Abstract

The present invention provides polynucleotides and polypeptides encoded thereby of human Type 2 RNase H. Methods of using these polynucleotides and polypeptides in enhancing antisense oligonucleotide therapies are also provided.

What is claimed is:  
     
         1 . An isolated polynucleotide encoding the human Type 2 RNase H polypeptide encoded by the nucleic acid sequence of the cDNA contained within ATCC Deposit No. 98536.  
     
     
         2 . A vector comprising a nucleic acid encoding the human Type 2 RNase H polypeptide encoded by the nucleic acid sequence of the cDNA contained within ATCC Deposit No. 98536.  
     
     
         3 . A host cell comprising the vector of  claim 2 .  
     
     
         4 . A composition comprising a vector comprising a nucleic acid encoding the human Type 2 RNase H polypeptide encoded by the nucleic acid sequence of the cDNA contained within ATCC Deposit No. 98536.  
     
     
         5 . The composition of  claim 4  further comprising an antisense oligonucleotide.  
     
     
         6 . A composition comprising an antisense oligonucleotide and a human Type 2 RNase H polypeptide, wherein the human Type 2 RNase H polypeptide is the human Type 2 RNase H polypeptide encoded by the nucleic acid sequence of the cDNA contained within ATCC Deposit No. 98536.  
     
     
         7 . A human Type 2 RNase H—his-tag fusion polypeptide, wherein the human Type 2 RNase H polypeptide is the human Type 2 RNase H polypeptide encoded by the nucleic acid sequence of the cDNA contained within ATCC Deposit No. 98536.  
     
     
         8 . A method of screening oligonucleotides to identify effective antisense oligonucleotides for inhibition of expression of a selected target protein comprising: 
 (a) contacting the human Type 2 RNase H polypeptide encoded by the nucleic acid sequence of the cDNA contained within ATCC Deposit No. 98536 with an RNA encoding the selected target protein and an oligonucleotide complementary to at least a portion of the RNA under conditions in which an RNA-oligonucleotide duplex is formed;    (b) detecting cleavage of the RNA of the RNA-oligonucleotide duplex wherein cleavage is indicative of antisense efficacy.    
     
     
         9 . The method of  claim 8  further comprising determining the site on the RNA at which cleavage occurs, whereby said site is identified as a Type 2 RNase H-sensitive site.  
     
     
         10 . The method of  claim 9  further comprising identifying an effective antisense oligonucleotide which hybridizes to said Type 2 RNase H-sensitive site.  
     
     
         11 . The method of  claim 8  wherein the oligonucleotide is one of a mixture or library of oligonucleotides.  
     
     
         12 . A method of identifying agents which increase or decrease activity or levels of the human Type 2 RNase H polypeptide encoded by the nucleic acid sequence of the cDNA contained within ATCC Deposit No. 98536 in a host cell comprising: 
 (a) contacting a cell in vitro expressing the human Type 2 RNase H polypeptide with an agent suspected of increasing or decreasing activity or levels of the human RNase H polypeptide; and    (b) measuring the activity or levels of the human RNase H polypeptide in the presence and absence of the agent so that an increase or decrease in the activity or levels of the human RNase H polypeptide can be determined.