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Publication Date

2024-5

First Advisor

Steven A. Williams

Document Type

Honors Project

Degree Name

Bachelor of Arts

Department

Biochemistry

Keywords

genetics, biochemistry, microbiology, immunology, mosquito, CRISPR-Cas9, gene editing, gene drive, JAK-STAT, PIAS

Abstract

Neglected Tropical Diseases (NTDs) present a considerable public health threat to more than one billion people worldwide – especially to communities with high rates of poverty, little access to medical care, and inadequate waste disposal methods. Aedes albopictus, the Asian tiger mosquito, is one of the main vectors for several diseases – most notably, the dengue, chikungunya, and Zika viruses. Due to the prevalence of Ae. albopictus and other such vectors, methods to control the spread of disease are essential to eliminating the threat of NTDs. This research aims to study the impacts of a CRISPR-Cas9-mediated gene knock-out of the PIAS negative regulator in the JAK-STAT immune pathway. We hypothesize that the knock-out of PIAS will upregulate multiple genes in the immune system and the production of antimicrobial peptides (AMPs). The work presented here reports on the initial efforts to transfect an Ae. albopictus C6/36 cell line with a CRISPR-Cas9 plasmid designed to knock out PIAS, and the preparation to perform RT-qPCR comparative gene analysis on two immune genes hypothesized to be directly impacted by this knock-out. Additionally, this paper describes the challenges encountered and future directions for completing this project.

Rights

©2024 Jane Dawson. Access limited to the Smith College community and other researchers while on campus. Smith College community members also may access from off-campus using a Smith College log-in. Other off-campus researchers may request a copy through Interlibrary Loan for personal use.

Language

English

Comments

67 pages: color illustrations. Includes bibliographical references.

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