Artificial riboswitch for controlling pre-mRNA splicing

Abstract

The present invention relates to riboswitches that have been engineered to regulate pre-mRNA splicing. In particular, the insertion of a high affinity theophylline binding aptamer into the 3′ splice site region, 5′ splice site region, or branchpoint sequence (BPS) of a pre-mRNA modulates RNA splicing in the presence of theophylline. Accordingly, the aspects of the present invention include, but are not limited to, theophylline-dependent riboswitches which modulate RNA splicing, methods of modulating RNA splicing using theophylline and its corresponding riboswitches, methods of improving/identifying theophylline-dependent riboswitches, methods of treating diseases associated with or caused by abnormal RNA splicing.

1. An artificial riboswitch comprising a nucleic acid sequence set forth as SEQ ID NO: 6. 
     
     
       2. A nucleic acid comprising the complementary sequence of the nucleic acid sequence in  claim 1 . 
     
     
       3. A vector comprising the nucleic acid of  claim 2 . 
     
     
       4. A cell comprising the artificial riboswitch of  claim 1  or the nucleic acid of  claim 1  or  2 . 
     
     
       5. A ligand-dependent riboswitch comprising a theophylline dependent aptamer, wherein the theophylline dependent aptamer is associated with the branchpoint sequence-3′ splice region in a pre-mRNA in a nucleic acid sequence set forth as, and the presence of theophylline modulates pre-mRNA splicing. 
     
     
       6. An isolated cell comprising the artificial riboswitch of claim 1 or the nucleic acid of claim 1 or 2.