ALTERATION OF NEURONAL GENE EXPRESSION BY SYNTHETIC piRNAs AND BY ALTERATION OF piRNA FUNCTION

Abstract

Provided herein are compositions and methods for the alteration of neuronal methylation by synthetic piRNAs or by alteration of piRNA function. Such alterations find use in the regulation and control of neural gene expression and concomitant neural functions. Further provided herein are systems and methods for the identification of target sites for regulation by piRNAs.

We claim: 
     
         1 . A method for altering neural gene expression, comprising: administering a synthetic piRNA to a subject under conditions such that methylation of a regulatory sequence regulating said gene is altered thereby altering said expression of said neural gene. 
     
     
         2 . The method of  claim 1 , wherein said altered methylation of said regulatory sequence of said gene is detected by methylome sequencing. 
     
     
         3 . The method of  claim 1 , wherein said methylome sequencing comprises sodium bisulfate conversion. 
     
     
         4 . The method of  claim 1 , wherein said methylome sequencing comprises methylome sequencing of promoter and gene body regions. 
     
     
         5 . The method of  claim 1 , further comprising strand-specific mRNA and/or smRNA sequencing. 
     
     
         6 . The method of  claim 1 , wherein said gene expression regulates a neurological function. 
     
     
         7 . The method of  claim 6 , wherein said neurological function is memory. 
     
     
         8 . The method of  claim 6 , wherein said neurological function is learning. 
     
     
         9 . The method of  claim 6 , wherein said neurological function is a pathology. 
     
     
         10 . The method of  claim 9 , wherein said pathology is a traumatic brain injury, a psychiatric disease, a cognitive disease, a neurodegenerative disease, or a post-traumatic stress disorder (PTSD). 
     
     
         11 . The method of  claim 1 , wherein said synthetic piRNA silences said neural target gene expression. 
     
     
         12 . The method of  claim 1 , wherein said synthetic piRNA molecule comprises a chemical modification that improves nuclease stability, decreases likelihood of triggering an innate immune response, lowers the incidence of off-target effects, or improves pharmacodynamics relative to a non-modified piRNA. 
     
     
         13 . The method of  claim 1 , wherein said synthetic piRNA comprises a nucleotide with at least one chemical modification selected from the group consisting of a phosphorothioate, a boranophosphate, a 4′-thio-ribose, a locked nucleic acid, a 2′-O-(2′-methoxyethyl), a 2′-O-methyl, a 2′-fluoro, a 2′-deoxy-2′-fluoro-b-D-arabinonucleic acid, a Morpholino nucleic acid analog, and a Peptide nucleic acid analog. 
     
     
         14 . The method of  claim 1 , wherein said synthetic piRNA is attached to a nanoparticle configured to cross the blood-brain barrier.