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Author

Eric Jedel

Date of Award

2019

Document Type

Restricted Thesis: Campus only access

Degree Name

Bachelor of Science

Department

Biology

Abstract

The ability to eliminate gene expression in organisms is useful for microbiologists to study genefunction, especially when investigating genes vital for survival. CRISPR interference (CRISPRi) has been shown to be an effective method of knocking down transcription of specific genes in model organisms. This thesis details a set of experiments to develop CRISPRi as a tool for gene knockdown in Sodalis glossinidius, a non-model bacteria endosymbiont of the tsetse fly. First, CRISPRi was shown to be effective for knocking down expression of acsD, a gene required for the production of iron-sequestering siderophores, as evidenced by qPCR and phenotypic confirmation. Then, the tool was used to target transcription of chiA, which encodes a chitinase that is predicted to degrade chitin from the fly gut into N-Acetyl D-Glucosamine (NAG) for S. glossinidiusmetabolism. Reduction of chiAexpression is relevant for metabolomic studies of the organism as well as investigating potential impacts NAG might have for increasing tsetse fly susceptibility to Trypanosoma brucei, the causative parasite for Human African Trypanosomiasis (HAT).

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