CRISPR/Cas9 mediated sex-ratio distortion by sex specific gene editing in Aedes aegypti

Aedes aegypti is a principal vector for several viruses including dengue virus, chikungunya virus and zika virus. Economic burden of mosquito-borne diseases, relative failure of traditional control strategies and the resistance development against insecticides enforces towards genetic manipulation of Ae. aegypti. Hence, a key gene doublesex (Aedsx) which regulate sex differentiation and alternatively splices to form male and female specific transcripts (Aedsx(M) and Aedsx(F)). CRISPR/Cas9 technique was employed to sex specifically disrupt the female-specific isoforms, Aedsx(F1) and Aedsx(F2), both of which were shown to be expressed only in female mosquitoes. Targeting of dsx(F) at the developmental stage has resulted in various phenotypic anomalies of adult females. The rate of adult mutation phenotype was recorded between 29 and 37% along with anomalies of wing size, proboscis length and reduction in the sizes of pre-blood-meal and after blood-meal ovaries in dsx(F1) and dsx(F2) microinjected groups, respectively. These findings can be correlated with reduced fecundity rate of G(o) female, where Aedsx(F1) and Aedsx(F2) groups showed reduction rate in range of 23–31%. Furthermore, hatching inhibition rate of 28 to 36% was also observed in G1 generation when compared to the wildtype. Overall, these results demonstrated that Aedsx(F) disruption has resulted in multiple female traits disruption including decreased fertility of the female that could directly or indirectly associated with reproduction and its disease transmitting abilities. All these findings suggesting that CRISPR works to alter the developmental pathways as predicted, and therefore this method potentially gives us the basis for the sex-ratio distortion system as genetic control approach for the management of this vector.