Polymearase III-based expression of therapeutic RNAs

Abstract

A transcribed non-naturally occurring RNA molecule comprising a desired 10 RNA molecule, wherein the 3+ region of the RNA is able to base-pair with at least 8 bases at the 5′ terminus of the same RNA molecule.

1 . A transcribed non-naturally occurring RNA molecule having hairpin structure, comprising a desired RNA portion, wherein said desired RNA portion is present between a 3′ region and 5′ region, wherein said 3′ region and said 5′ region form an intramolecular stem with each other comprising at least 8 base pairs.  
     
     
         2 . The RNA molecule of  claim 1 , wherein said RNA molecule is transcribed by a RNA polymerase III based promoter system.  
     
     
         3 . The RNA molecule of  claim 1 , wherein said RNA molecule is transcribed by a type 2 pol III promoter system.  
     
     
         4 . The RNA molecule of  claim 1 , wherein said RNA molecule is a chimeric tRNA.  
     
     
         5 . The RNA molecule of  claim 3 , wherein said RNA molecule has A and B boxes of a type 2 pol III promoter separated by between 0 and 300 bases.  
     
     
         6 . The RNA molecule of  claim 5 , wherein said desired RNA portion is at the 3′ end of said B box of said RNA molecule.  
     
     
         7 . The RNA molecule of  claim 5 , wherein said desired RNA portion is in between said A and said B box of said RNA molecule.  
     
     
         8 . The RNA molecule of  claim 5 , wherein said desired RNA portion includes said B box of said RNA molecule.  
     
     
         9 . The RNA molecule of  claim 1 , wherein said desired RNA portion is selected from the group consisting of antisense RNA, decoy RNA, therapeutic editing RNA, enzymatic RNA, agonist RNA and antagonist RNA.  
     
     
         10 . The RNA molecule of  claim 1 , wherein said 5′ region of said RNA molecule is able to base-pair with at least 12 bases of said 3′ region.  
     
     
         11 . The RNA molecule of  claim 1 , wherein the 5′ region of said RNA molecule is able to base-pair with at least 15 bases of said 3′ region.  
     
     
         12 . A DNA vector encoding the RNA molecule of  claim 1 .  
     
     
         13 . A RNA vector encoding the RNA molecule of  claim 1 .  
     
     
         14 . The DNA vector of  claim 12  wherein the portions of the DNA vector encoding said RNA molecule function as a RNA pol III promoter.  
     
     
         15 . A cell comprising the vector of  claim 12 .  
     
     
         16 . A cell comprising the vector of  claim 13 .  
     
     
         17 . A cell comprising the RNA of  claim 1 .  
     
     
         18 . A method to provide a desired first RNA molecule having hairpin structure in a cell, comprising introducing into said cell a second RNA molecule comprising a 5′ region, a 3′ region, and said desired first RNA molecule, wherein said 5′ terminus is able to base pair with at least 8 bases of said 3′ region, and wherein said desired first RNA molecule is present between the bases of the 3′ region and the 5′ terminus capable of base pairing in the second RNA molecule under conditions suitable to provide the desired first RNA molecule in the cell.  
     
     
         19 . The method of  claim 18 , wherein the introduction of the second RNA molecule comprises providing a vector encoding said second RNA molecule.  
     
     
         20 . The RNA molecule of  claim 1 , wherein said RNA molecule is transcribed by a RNA polymerase II promoter system.  
     
     
         21 . The RNA molecule of  claim 1 , wherein said RNA molecule is transcribed by a U6 small nuclear RNA promoter system.  
     
     
         22 . The RNA molecule of  claim 1 , wherein said RNA molecule is transcribed by an adenovirus VA1 RNA promoter system.  
     
     
         23 . The RNA molecule of  claim 1 , wherein said RNA molecule is a chimeric adenovirus VA1 RNA.  
     
     
         24 . The RNA molecule of  claim 1 , wherein said intramolecular stem is separated from said desired RNA portion by spacer sequence.  
     
     
         25 . The RNA molecule of  claim 24 , wherein said spacer sequence is about 5-50 nucleotides.