*Please note that this project can be completed as a PhD or MPhil*
The process of sperm production in animals requires activation of many genes that are not used in any other tissues. Indeed, more than 10% of the protein coding genes in both the human and fly genomes are expressed exclusively in the testis. In our lab we investigate the transcriptional controls that generate this testis specificity – both keeping the genes off in other tissues and (the main focus of the lab) activating them in spermatocytes in testes.
A critical, conserved, transcriptional regulatory complex activates expression of many genes in spermatocytes in Drosophila melanogaster. We have mutant alleles of most of the components and have detailed phenotypic analysis, including for example RNAseq to identify transcriptional changes. We have additionally identified a putative novel component of the complex, via protein-interaction assays with known complex subunits. The protein localizes to chromatin in the male germline, consistent with a role in gene expression. We are using CRISPR mutagenesis to generate knock out alleles of this gene. This project will involve analysis of the mutants, initially aiming to determine how the phenotype compares to known pathway components. Further analysis will reveal how the protein interacts with, and modulates the function of, the core complex. The gene has recently evolved, and a cross species comparison will reveal how new components can integrate into established regulatory modules.
Molecular biology – DNA and RNA extraction, PCR, RT-PCR, Q-RT-PCR. Plasmid construction.
DNA-protein interaction analysis – ChIP seq.
RNA seq – sample preparation, library construction, bioinformatics.
Cytology and microscopy of testes; immunostaining; RNA in situ hybridisation.
Drosophila genetics, including CRISPR tagging, transgenesis and inducible expression systems.