Construct of RNAi plasmids and Their Application For the Study of Meiotic Proteins Through RNA Interference

Sexual reproduction in eukaryotes requires production of haploid gametes from diploid cells. This is accomplished through meiosis, in which two successive divisions of a diploid cell in the germ line results in four haploid gametes. This process is highly regulated, and requires the action of many key proteins in order for chromosomes to segregate properly and meiosis to be successful. In the absence of a protein involved in meiosis, one may expect chromosome missegregation, failure of meiosis, and eventual cell apoptosis. In this study, the function of meiotic proteins are analyzed through the process of RNA interference in Caenorhabditis elegans, a model organism for the study of meiosis. RNA interference (RNAi) reversibly modifies the expression of proteins due to the introduction of double‐stranded RNA (dsRNA), which results in the reversible depletion, or silencing, of the protein encoded by the complementary gene. In previous work, we used proteomics to identify candidate meiosis proteins. In order to evaluate these candidates, we constructed RNAi plasmids that would target the gene under investigation. A successful RNAi construct will result in the decreased expression of the target protein. If the protein is required for proper completion of meiosis, the process will be disrupted, resulting in the reduction in viability of C. elegans progeny. This poster will present data on the construct of RNAi plasmids and their subsequent effects on meiosis in C. elegans , as well as direct analysis of the cytology of meiosis through fluorescence microscopy.