Investigating baculovirus as a viral vector for delivery of Cas9 to the ovaries of disease vector Aedes aegypti for use in mosquito gene editing technologies
Open Access
Author:
Mckeand, Sage
Area of Honors:
Elective Area of Honors - Entomology
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Jason Laurence Rasgon, Thesis Supervisor Gary Felton, Thesis Honors Advisor
Keywords:
entomology vector control disease pathogen baculovirus AcMNPV viral vector vector Aedes aegypti gene editing Cas9 ReMOT Control mosquito ovaries
Abstract:
Genetic engineering is an emerging tool for reducing the transmission of pathogens such
as Zika, dengue, and West Nile viruses from the insect vector Aedes aegypti to humans. Recent
developments in specific gene modification techniques have led to more widely accessible
methods of editing, but with similarly low efficiencies to traditional gene editing protocols. By
using the baculovirus AcMNPV as a vector for Cas9, I aim to increase the efficiency of targeted
gene modification in Ae. aegypti. This manuscript details initial experiments to understand how a
baculovirus vector expressing a fluorescent protein gene (mCherry) will enter the ovaries of a
female adult mosquito. Based on imaging, it appears that the baculovirus delivers synthetic DNA
encoding mCherry into the follicle of the oocyte. This opens possibilities for further experiments
including using an ovary specific localization signal to direct Cas9 expressed from the AcMNPV
genome for targeted mutagenesis in mosquitoes.