High-throughput RNAi screening for germline apoptosis genes in Caenorhabditis elegans.

Among the greatest tools that Caenorhabditis elegans can provide researchers are the capabilities to perform high-throughput, genome-wide screens. Using bacterial RNAi libraries, which cover the majority (>85%) of the worm genome, genes can be rapidly and systematically evaluated for apoptosis phenotypes in the germline. Screens can be designed to directly assess the levels of apoptotic corpses under normal physiological conditions using transgenic strains expressing fluorescent reporters that mark apoptotic bodies. Vital dyes that are selectively retained in apoptotic cells, such as acridine orange (AO), can also be used to screen for genes that regulate germline apoptosis. Using these reagents, screens can be performed in wild-type worms or mutant backgrounds that suppress or enhance apoptosis phenotypes. This protocol describes methods for designing and carrying out high-throughput or targeted RNAi screens for germline apoptosis regulators.