425A Poster - 05. Reproduction and gametogenesis
Thursday April 07, 2:00 PM - 4:00 PM

Authors:
William Gilliland; Amanda Bowen ; Kelly Conger; Doreen Elrad; Marcin Marciniak; Denny May; Gabrielle Presbitero

Affiliation: DePaul University

Keywords:
b. oogenesis; c. RNAi

Forward genetic screens induce mutations, make the mutated chromosomes homozygous, and then homozygotes for the phenotype of interest. When studying female meiosis, the phenotype is usually nondisjunction from chromosome segregation errors. This means that mutant females must be viable and fertile, and any meiotic genes that are lethal or sterile when homozygous cannot be recovered by this approach. Our lab has screened the VALIUM22 collection produced by the Harvard TRiP Project, which contains RNAi constructs targeting genes known to be expressed in the germline in a vector optimized for germline expression. By driving RNAi with GAL4 under control of a germline-specific promoter (nos or mat-alpha), we can test genes that would be lethal if knocked down in all cells, and by examining unfertilized metaphase-arrested mature oocytes, we can identify defects associated with genes whose knockdown results in sterility.
We screened this collection to identify genes that disrupt either of two phenotypes: the ability of meiotic chromosomes to congress to a single mass at the end of prometaphase, and the sequestration of Mps1-GFP to unknown structural filaments in response to hypoxia. After screening the ~1500 lines in the collection, we obtained multiple hits for both phenotypes, identified novel meiotic phenotypes for genes that had been previously characterized in other tissues, and found novel phenotypes for several previously uncharacterized genes.