RNA interference pathway genes as tools for targeted genetic interference

Abstract

Genes involved in double-stranded RNA interference (RNAi pathway genes) are identified and used to investigate the RNAi pathway. The genes and their products are also useful for modulating RNAi pathway activity.

1 . A method of inhibiting the activity of a gene, the method comprising introducing an RNAi agent into a cell, wherein the RNAi agent is prepared by incubating a double-stranded RNA (dsRNA) targeted to the gene in the presence of an RNAi pathway component, wherein the RNAi component is an RDE-1 polypeptide or homolog thereof.  
     
     
         2 . The method of  claim 1 , wherein the RNAi pathway component is an RDE-1 polypeptide.  
     
     
         3 . A method of inhibiting the activity of a gene, the method comprising introducing an RNAi agent into a cell, wherein the RNAi agent is prepared by incubating a double-stranded RNA (dsRNA) targeted to the gene in the presence of an RNAi pathway component, wherein the RNAi component is an RDE-4 polypeptide or homolog thereof.  
     
     
         4 . The method of  claim 3 , wherein the RNAi pathway component is an RDE-4 polypeptide.  
     
     
         5 . A method of inhibiting the activity of a gene, the method comprising introducing an RNAi agent into a cell, wherein the RNAi agent is prepared by incubating a dsRNA in the presence of at least a first and second RNAi pathway component, wherein the first RNAi component is an RDE-1 polypeptide or homolog thereof and the second RNAi component is an RDE-4 polypeptide or homolog thereof.  
     
     
         6 . The method of  claim 5 , wherein the RNAi pathway components are an RDE-1 polypeptide and an RDE-4 polypeptide.  
     
     
         7 . The method of  claim 1 ,  3  or  5 , wherein the RNAi agent is introduced into the cell in a liposome.  
     
     
         8 . The method of  claim 1 ,  3  or  5 , wherein the RNAi agent is introduced into the cell by injection.  
     
     
         9 . The method of  claim 1 ,  3  or  5 , wherein the cell is in an animal.  
     
     
         10 . The method of  claim 2  or  6 , wherein the RDE-1 polypeptide comprises an amino acid sequence at least 80% identical to the amino acid sequence set forth as SEQ ID NO:3 and is capable of promoting RNAi.  
     
     
         11 . The method of  claim 10 , wherein the RDE-1 polypeptide comprises an amino acid sequence at least 95% identical to the amino acid sequence set forth as SEQ ID NO:3.  
     
     
         12 . The method of  claim 10 , wherein the RDE-1 polypeptide comprises the amino acid sequence set forth as SEQ ID NO:3.  
     
     
         13 . The method of  claim 4  or  6 , wherein the RDE-4 polypeptide comprises an amino acid sequence at least 80% identical to the amino acid sequence set forth as SEQ ID NO:5 and is capable of promoting RNAi.  
     
     
         14 . The method of  claim 13 , wherein the RDE-4 polypeptide comprises an amino acid sequence at least 95% identical to the RDE-4 amino acid sequence set forth as SEQ ID NO:5.  
     
     
         15 . The method of  claim 13 , wherein the RDE-4 polypeptide comprises the RDE-4 amino acid sequence set forth as SEQ ID NO:5.