LAUSR

Significance Here, we describe the generation of Aedes aegypti mosquitoes that are engineered to be resistant to Zika virus (ZIKV) transmission. Our results demonstrate that engineered mosquitoes express a polycistronic cluster of synthetic small RNAs designed to target the ZIKV genome. As a result, homozygous mosquitoes were refractory to ZIKV infection, and therefore could not transmit the virus. Additionally, mosquitoes heterozygous for the transgene showed significantly lower levels of viral infection, dissemination, and transmission compared with wild-type mosquitoes; importantly, these levels were low enough to make such mosquitoes unlikely to transmit ZIKV to a susceptible host. Finally, we discuss how such an engineering approach can be used to combat the major health burden of ZIKV, and potentially other arboviruses, in the future. Recent Zika virus (ZIKV) outbreaks have highlighted the necessity for development of novel vector control strategies to combat arboviral transmission, including genetic versions of the sterile insect technique, artificial infection with Wolbachia to reduce population size and/or vectoring competency, and gene drive-based methods. Here, we describe the development of mosquitoes synthetically engineered to impede vector competence to ZIKV. We demonstrate that a polycistronic cluster of engineered synthetic small RNAs targeting ZIKV is expressed and fully processed in Aedes aegypti, ensuring the formation of mature synthetic small RNAs in the midgut where ZIKV resides in the early stages of infection. Critically, we demonstrate that engineered Ae. aegypti mosquitoes harboring the anti-ZIKV transgene have significantly reduced viral infection, dissemination, and transmission rates of ZIKV. Taken together, these compelling results provide a promising path forward for development of effective genetic-based ZIKV control strategies, which could potentially be extended to curtail other arboviruses.