The interaction between RNAs and RNA-binding proteins (RBPs) is an important topic in studies of gene expression. Our lab is interested in fbf-1, a gene that encodes an RBP that maintains stem cell proliferation and differentiation, and rrf-1, a gene that encodes an RNA-polymerase that generates small regulatory RNAs. These RNAs have been proposed to contribute to the function of FBF genes, but this hypothesis remains controversial in the field. Through previous lab research, we suspected that at 24°C, a strain of Caenorhabditis elegans (C. elegans) with rrf-1 and fbf-1 mutations becomes sterile over the course of multiple generations. Due to this finding, we decided to investigate if the rate at which rrf-1 and fbf-1 mutant C. elegans become sterile at 24°C changes based on how the mutant is produced. We use the model organism C. elegans because many of the proteins present in C. elegans have mammalian orthologs. We will generate the mutant strain in two ways. The first method is to produce a strain that has a fbf-1 deletion but maintains a wild type (WT) copy of rrf-1 through the use of a genetic balancer, a genetic tool that stabilizes a WT gene copy in heterozygotes. We hypothesize that the rrf-1 and fbf-1 mutant progeny of this strain will become sterile at over time at 24°C because the RRF-1-generated small RNAs they have inherited will be depleted each generation. The second method is to produce a strain that has an rrf-1 deletion but maintains a WT copy of fbf-1 using a genetic balancer. We hypothesize that the rrf-1 and fbf-1 mutant progeny will become sterile at 24°C immediately because they did not inherit RRF-1-generated small RNAs. Understanding the link between small regulatory RNAs and RBPs is important because their interaction is implicated in many human diseases, including cancer
