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