Meeting Abstract

P1.144  Saturday, Jan. 4 15:30  Establishing RNAi in Ciona intestinalis to determine gene function during peripheral nervous system development HURLESS, V.H.*; ZELLER, R.W.; San Diego State University; San Diego State University vhurless@mail.sdsu.edu

RNA interference, or RNAi, is a post-transcriptional process that regulates gene expression by silencing mRNA targets through degradation. RNA-induced silencing is mediated by base pairing of small RNA molecules, such as microRNAs, to their endogenous mRNA targets. Many studies incorporate the use of artificial microRNAs or siRNAs to silence genes to determine their function. Although, RNAi has been established in numerous model organisms, RNAi approaches have not yet been adapted and implemented in the marine invertebrate chordate Ciona intestinalis. Our lab has developed an innovative method to develop mRNA target-specific RNAi utilizing artificial microRNAs to knockdown genes of interest in C. intestinalis. Using this method, we have been successful in knocking down two genes that result in expected phenotypes, one of which displays a similar phenotype to a published null mutant. Refining this tool will allow for the widespread use of RNAi in C. intestinalis further demonstrating they are an ideal model for chordate development and genomics. In addition to establishing effective RNAi in C. intestinalis, my project goal is to utilize this tool to explore the epistatic relationship of regulatory genes involved in peripheral nervous system (PNS) development. Our lab has defined a gene regulatory network governing larval PNS sensory neurons and a Pou4 gene target list from RNA-sequencing data. Employing RNAi to knockdown these regulatory genes will elucidate their function and refine the PNS gene network.