Novel Assay System and Analysis Algorithm in Detecting RNA Editing Events and Linked Splicing Isoforms

Period of Performance: 07/01/2016 - 06/30/2017

$150K

Phase 1 STTR

Recipient Firm

Celetrix, LLC
MANASSAS, VA 20110
Principal Investigator

Abstract

AbstractRNA editing is a process in which the genome-encoded information is altered in RNA. RNA editing is anefficient way to increase RNA complexity, thereby fine-tuning both gene function and dosage. Adenosine toInosine (A-to-I) editing is the most common type of RNA editing known in animals. The cellular machineryrecognizes inosine as guanosine, so A-to-I editing of codons and splicing signals directly modifies protein-coding gene function, whereas editing of microRNAs and their binding sites alter gene expression. The vastmajority of A-to-I editing, however, is detected in non-coding regions (e.g. Alu-repeats within introns and 3' or 5'untranslated regions), strongly suggesting a still unknown regulatory role of this cellular mechanism.Dysregulation of mRNA editing was implicated in several neurological diseases. In addition, we and othergroups showed that alterations in mRNA editing of one of the serotonin receptors (serotonin 2C receptor) isassociated with completed suicide, major depression and possibly substance use disorder (SUD). Currently,short read Sanger or Illumina sequencing are the mainstream tools in RNA editing studies. However, thesetools cannot detect the ?phase? of editing events; i.e., they cannot detect simultaneous editing events at two ormore sites which are situated further than 50-100bp from one another on the same mRNA molecule. Inaddition, the current tools, which focus on a small region in the vicinity of a given editing site, do not allow us tostudy RNA editing in the context of splicing. In the proposed project we aim to develop a state-of-the-art toolthat can be used by the entire scientific community. If we are successful, this tool will enable a simultaneousdetection and quantification of RNA editing and splicing isoforms in the brain hence allowing researchers tostudy RNA editing in the context of splicing. Moreover, this tool will enable determination of the haplotypes ofmRNA molecules with multiple editing sites. We anticipate that this tool will have a great commercial potentialand will facilitate research on RNA editing as one of the molecular mechanisms that is implicated in SUD.