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

Abstract RNA editing is a process in which the genome encoded information is altered in RNA RNA editing is an efficient way to increase RNA complexity thereby fine tuning both gene function and dosage Adenosine to Inosine A to I editing is the most common type of RNA editing known in animals The cellular machinery recognizes 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 vast majority of A to I editing however is detected in non coding regions e g Alu repeats within introns and andapos or andapos 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 other groups showed that alterations in mRNA editing of one of the serotonin receptors serotonin C receptor is associated 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 these tools cannot detect the phase of editing events i e they cannot detect simultaneous editing events at two or more sites which are situated further than bp from one another on the same mRNA molecule In addition the current tools which focus on a small region in the vicinity of a given editing site do not allow us to study RNA editing in the context of splicing In the proposed project we aim to develop a state of the art tool that can be used by the entire scientific community If we are successful this tool will enable a simultaneous detection and quantification of RNA editing and splicing isoforms in the brain hence allowing researchers to study RNA editing in the context of splicing Moreover this tool will enable determination of the haplotypes of mRNA molecules with multiple editing sites We anticipate that this tool will have a great commercial potential and will facilitate research on RNA editing as one of the molecular mechanisms that is implicated in SUD Narrative Dysregulation of mRNA editing was implicated in several neurological diseases however the current technology has limitations in detecting mRNA editing events Here we aim to develop a state of the art tool that enables a simultaneous detection and quantification of RNA editing and splicing isoforms in the brain hence allowing researchers to study RNA editing in the context of splicing We anticipate that this tool will have a great commercial potential and will facilitate research on RNA editing as one of the molecular mechanisms that is implicated in SUD