ENGINEERING CAENORHOBDITIS ELEGANS FOR PHOTOTAXIS

The organism Blastocladiella emersonii is an aquatic fungus that was found to use the second messenger cGMP and retinal for phototaxis in the zoospore phase of its life cycle. B. emersonii phototaxis is mediated by a fusion protein, RhoGC, made up of a microbial rhodopsin domain and a eukaryotic guanylyl cyclase catalytic domain. Our lab is interested in this unique protein to elucidate the mechanism by which light and retinal control activity of the guanylyl cyclase domain and to develop the protein as an optogenetic tool, extending its use in different laboratories as a technique to control cGMP levels in cells and organisms with light. Here we begin studies to express RhoGC in the AWB olfactory and AWC chemo/thermosensory neurons of Caenorhabditis elegans to test whether we can engineer this organism for phototaxis. We describe here construction of vectors for targeted expression of a RhoGC gene into C. elegans. We will next insert the finished vectors into C.elegans and test their viability for induced phototaxis.