The FAPESP-Keele network phase 2: generation of mutant Aedes aegypti lines using CRISPR-Cas9

Abstract

To deal with the risk of infection caused by exposure to various microbes, insects have developed an immune system compound with a variety of defense mechanisms working in synergy. Recent unpublished studies developed in our laboratory, under the FAPESP-Keele network, with Ae. aegypti population with different levels of susceptibility to DENV4 revealed that these populations have different transcriptional responses when infected with the virus. In order to perform functional testing on target genes, this proposal is intended to use a CRISPR-Cas9 genome-editing tool for the identification of restriction factors to dengue virus in Ae. aegypti. For such, various Ae. aegypti mutant strains (each gene chosen will be knocked out individually) will be generated in the platform of insect vectors transformation of the Centre of Applied Entomology and Parasitology (CAEP), Keele University, in collaboration with Dr. Julien Pelletier. The generated lines will be brought to IBTEC/UNESP, where the strains infection test with DENV will be conducted. (AU)