Understanding the Role of the Duplicated Gene D2005.1

Adenosine to inosine (A to I) RNA editing is a conserved process catalyzed by adenosine deaminases that act on dsRNA (ADARs). In mice, knockout of ADAR genes is lethal, Caenorhabditis elegans lacking A to I editing are viable. C. elegans possess two ADAR genes: adr-1 and adr-2. ADR-2 catalyzes the deamination, while ADR-1 acts as a regulator of ADR-2 activity. In C. elegans, RNA editing was shown to function in cellular immunity and both ADAR genes were shown to be involved. Our goal is to understand the mechanism of regulation of editing and to identify its players. One candidate regulator is D2005.1 an exact duplication of adr-1 C terminal region, Located only 6-7 kb apart from adr-1. Preliminary qPCR results show that partial deletion of D2005.1 leads to overexpression of adr-1. Unfortunately, the existing ADR-1 and D.2005.1 mutant strains possess partial deletions of the genes and thus do not allow to differentiate between transcripts originating from adr-1 and D2005.1 Therefore, to be able to study the functions of D.2005.1 we generated, using CRISPR, three new C. elegans stains a full deletion of adr-1 a full deletion of D2005.1 and a double mutant. We plan to use the mutants to: 1. elucidate the effect of D2005.1 deletion on the expression of the different isoforms of adr-1 using RTRT-PCR 2. perform expression analysis of the 3 strains by generating and sequencing directional RNA libraries. 3. check
how D2005.1 deletion affects different phenotypic traits of the worm.