Extranuclear RNA splicing in neuronal dendrites

Abstract

The present invention relates to methods of synaptic network remodeling by means of extranuclear RNA splicing. The present invention also provides methods of extranuclear RNA splicing, and methods of protein translation based on extranuclear RNA splicing.

1 . A method of remodeling a dendrite, said method comprising the steps of: 
 a. transfecting a dendrite with an RNA comprising at least one intron, wherein said dendrite comprises at least one component of a spliceosome and further wherein said component of a spliceosome is capable of splicing an RNA;    b. allowing said RNA comprising at least one intron to be spliced by said spliceosome components; and    c. allowing said spliced RNA to be translated in said dendrite;    wherein said dendrite is thereby remodeled as a consequence of said translation.    
     
     
         2 . A method of remodeling a dendrite interaction, said method comprising the steps of: 
 a. transfecting a dendrite with an RNA comprising at least one intron, wherein said dendrite comprises at least one component of a spliceosome and further wherein said component of a spliceosome is capable of splicing an RNA;    b. allowing said RNA comprising at least one intron to be spliced by said spliceosome components; and    c. allowing said spliced RNA to be translated in said dendrite;    wherein said dendrite interaction is thereby remodeled as a consequence of said translation.    
     
     
         3 . A method of remodeling a synaptic network comprising interaction with at least one dendrite, said method comprising the steps of: 
 a. transfecting a dendrite with an RNA comprising at least one intron, wherein said dendrite comprises at least one component of a spliceosome and further wherein said component of a spliceosome is capable of splicing an RNA;    b. allowing said RNA comprising at least one intron to be spliced by said spliceosome components; and    c. allowing said spliced RNA to be translated in said dendrite;    wherein said synaptic network is thereby remodeled as a consequence of said translation.    
     
     
         4 . A method of splicing an RNA, said method comprising the steps of: 
 a. providing an isolated dendrite comprising at least one component of a spliceosome, wherein said component is capable of splicing an RNA; and    b. transfecting said dendrite with an RNA comprising at least one intron;    wherein said RNA comprising at least one intron is spliced by said spliceosome components.    
     
     
         5 . The method of  claim 1 , wherein said dendrite is a component of a neuron.  
     
     
         6 . The method of  claim 1 , wherein said dendrite is an isolated dendrite.  
     
     
         7 . A method of splicing an RNA, said method comprising the steps of: 
 a. providing an isolated cell comprising at least one component of a spliceosome in the cytoplasm, wherein said component is capable of splicing an RNA; and    b. transfecting said cell with an RNA comprising at least one intron;    wherein said RNA comprising at least one intron is spliced by said spliceosome components in said cytoplasm.    
     
     
         8 . The method of  claim 1 , wherein RNA splicing donor/acceptor pairs are selected from the group consisting of canonical, atypical and cryptic.  
     
     
         9 . The method of  claim 1 , wherein said RNA comprising at least one intron is a pre-RNA.  
     
     
         10 . The method of  claim 9 , wherein said pre-RNA is a pre-mRNA.  
     
     
         11 . The method of  claim 1 , wherein said RNA comprising at least one intron is derived from a nucleic acid comprising a construct selected from the group consisting of pEGFP-N1, pDsRed-N1, SF1/mBBP-GFP, U2AF65-GFP, a GFP construct, a DsRed construct, a histone 2B-YFP construct.  
     
     
         12 . The method of  claim 1 , wherein at least one spliceosome component is selected from the group consisting of Y14, Magoh, RNPS 1, SC-35, SF2, U2AF65, Smith antigen, pan-SR antigen, U1 snRNP, U2 snRNP, U4 snRNP, U5 snRNP, U6 snRNP.  
     
     
         13 . A method of translating an RNA, said method comprising the steps of: 
 a. providing an isolated dendrite comprising at least one component of a spliceosome, wherein said component is capable of splicing an RNA; and    b. transfecting said dendrite with an RNA comprising at least one intron, wherein said RNA is spliced by said spliceosome components;    further wherein said spliced RNA is translated.    
     
     
         14 . A method of translating an RNA, said method comprising the steps of: 
 a. providing an isolated cell comprising at least one component of a spliceosome in the cytoplasm, wherein said component is capable of splicing an RNA; and    b. transfecting said cell with an RNA comprising at least one intron, wherein said RNA is spliced by said spliceosome components in said cytoplasm;    further wherein said spliced RNA is translated.    
     
     
         15 . A method of splicing an RNA, said method comprising the steps of: 
 a. providing an isolated synaptoneurosome comprising at least one component of a spliceosome, wherein said component is capable of splicing an RNA; and    b. contacting said synaptoneurosome with an RNA comprising at least one intron;    wherein said RNA comprising at least one intron is spliced by said spliceosome components.    
     
     
         16 . A method of translating an RNA, said method comprising the steps of: 
 a. providing an isolated synaptoneurosome comprising at least one component of a spliceosome, wherein said component is capable of splicing an RNA;    b. contacting said synaptoneurosome with an RNA comprising at least one intron, wherein said RNA is spliced by said spliceosome components; and    c. contacting said spliced RNA with a composition capable of translating an RNA under conditions suitable for translating an RNA.