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Research Project: Using Transcriptomics and Proteomics to Understand Arbovirus Effects on Midge Sensory Function

Location: OPERATIONS

Project Number: 3022-32000-012-005-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Aug 1, 2020
End Date: Jul 31, 2025

Objective:
This project will use bioinformatic and molecular approaches to assess effects of orbivirus infection on sensory gene expression in female Culicoides sonorensis.

Approach:
RNA and protein extracted from midges who were infected with orbiviruses (Bluetongue virus, Epizootic Hemorrhagic Disease Virus) and Vesicular Stomatitis Virus via microinjection and/or feeding will be compared to controls (blood fed, un-injected, sham-injected). To determine differences in gene and protein expression across tissues and treatments, the graduate student and Cooperator will perform library/sample preparation, normalization, sequencing and bioinformatic analyses of the data generated from these experiments, including de novo assembly of reads, functional annotation, differential gene expression, GO enrichment, KEGG pathway mapping, tandem mass tagging (TMT), LC-MS (for protein expression analysis) and other techniques deemed appropriate to the study. Differentially-expressed transcripts and proteins/peptides will be confirmed by qRT-PCR and western analysis by others on the team. The overarching goal for this project is to pair these gene and protein expression phenotypes with behavioral phenotypes, such as those measured by electropenetrogram, electrophysiology (photoreceptor and antennae function) and behavioral assays (attraction or repulsion to light wavelengths and olfactory/gustatory cues).