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ViewAttendees 18

Member Symposium

Arthropod Saliva: Role in Blood Feeding and Pathogen Transmission

3: Towards a functional characterization of mosquito salivary proteins through CRISPR/Cas9 gene editing techniques

On-Demand
On-Demand
Mosquitoes saliva facilitates blood meal acquisition through pharmacologically active compounds that prevent host hemostasis and immune responses. Vasodilators, anticoagulants and platelet aggregation-inhibitors are present in insect’s saliva to counteract the three branches of hemostasis and ensure a successful blood-feeding. We generated  homozygous knock out (KO) mosquito lines by CRISPR/Cas9 system to further characterize the biological function of Sialokinin and two D7 salivary proteins by loss-of-function studies. Sialokinin, a vasodilatory decapeptide, related to the tachykinin family acts by stimulating endothelial cells via cGMP to produce nitric oxide. The D7s, one of the most abundant salivary proteins act as scavengers of biogenic amines and eicosanoids involved in hemostasis. All KO mosquito lines took longer than parental wild type mosquitoes to locate blood vessels when feeding on mice and, therefore, had significantly longer probing times. In D7 KO mosquitoes, the differences in probing time were abolished when B6.129S2-Alox5tm1Fun mutant mice were used. These mutant mice do not produce leukotrienes, confirming the role of D7 proteins as leukotrienes scavengers. We also investigated the role of D7 salivary proteins in Plasmodium gallinaceum infection. Our results show that both KO lines carried significantly less oocysts per midgut and reduction in sporozoites number in salivary glands. The information generated by this work highlights the biological functionality of salivary gene products in blood feeding and pathogen transmission.