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OSR Journal of Student Research

Article Title

Anatomical Locations of the dop-1 Expression Sites in C. elegans

Abstract

Our laboratory utilizes the model organism Caenorhabditis elegans to study the molecular basis of behavior. We and others have demonstrated that behavioral avoidance of noxious chemicals in C. elegans is modulated by the catecholamine dopamine (DA). To further investigate the details of these mechanisms, our project uses CRISPR-Cas9 as a means of conducting gene-substitution experiments. Fluorescent reporter genes replace the coding sequence of the dop-1 dopamine receptor gene; this visibly reveals the precise anatomical locations of their expression sites. This is accomplished by engineering two DNA plasmids. One plasmid contains both the Cas9 and guide RNA sequences, which we will generate by site-directed mutagenesis. The other plasmid provides a repair template containing the fluorescent reporter sequence flanked by the appropriate genomic DNA, created by Gibson assembly. These constructs will be injected in C. elegans germline cells, and animals carrying modified genomes will be screened for using fluorescence microscopy and polymerase chain reaction. We will report on our progress creating the recombinant plasmids.

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