Faculty, Staff and Student Publications
Publication Date
7-17-2024
Journal
BMC Genomics
Abstract
Due to limitations in conventional disease vector control strategies including the rise of insecticide resistance in natural populations of mosquitoes, genetic control strategies using CRISPR gene drive systems have been under serious consideration. The identification of CRISPR target sites in mosquito populations is a key aspect for developing efficient genetic vector control strategies. While genome-wide Cas9 target sites have been explored in mosquitoes, a precise evaluation of target sites focused on coding sequence (CDS) is lacking. Additionally, target site polymorphisms have not been characterized for other nucleases such as Cas12a, which require a different DNA recognition site (PAM) and would expand the accessibility of mosquito genomes for genetic engineering. We undertook a comprehensive analysis of potential target sites for both Cas9 and Cas12a nucleases within the genomes of natural populations of Anopheles gambiae and Aedes aegypti from multiple continents. We demonstrate that using two nucleases increases the number of targets per gene. Also, we identified differences in nucleotide diversity between North American and African Aedes populations, impacting the abundance of good target sites with a minimal degree of polymorphisms that can affect the binding of gRNA. Lastly, we screened for gRNAs targeting sex-determination genes that could be widely applicable for developing field genetic control strategies. Overall, this work highlights the utility of employing both Cas9 and Cas12a nucleases and underscores the importance of designing universal genetic strategies adaptable to diverse mosquito populations.
Keywords
Animals, Anopheles, CRISPR-Cas Systems, Aedes, Genetic Variation, RNA, Guide, CRISPR-Cas Systems, Endodeoxyribonucleases, CRISPR-Associated Proteins, Genome, Insect, Mosquito Vectors, Gene Editing, Bacterial Proteins
Included in
Bioinformatics Commons, Biomedical Informatics Commons, Genomics Commons, Medical Genetics Commons, Oncology Commons
Comments
Supplementary Materials
Data Availability Statement
PMID: 39020310