Engineered resistance to Zika virus in transgenic Aedes aegypti expressing a polycistronic cluster of synthetic small RNAs
Abstract
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.
Additional Information
© 2019 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND). Edited by Alexander S. Raikhel, University of California, Riverside, CA, and approved December 31, 2018 (received for review June 22, 2018) We thank Prof. Robert Tesh and Dr. Nikos Vasilakis (University of Texas Medical Branch) for providing the ZIKV FSS13025 and PRVABC59 strains and Lee Trinidad for preparing viral stocks. We thank Christian Bowman for helping with the inverse PCR assay. We also thank Prof. Scott O'Neill (Institute of Vector-Borne Diseases, Monash University) and the World Mosquito Program for providing Wolbachia-infected mosquito eggs. Finally, we thank Dr. Guann-Yi Yu (National Tsing Hua University) for providing the Stat^(−/−) mice used in this study. This work was supported, in part, by an NIH-K22 Career Transition Award (5K22AI113060), an NIH Exploratory/Developmental Research Grant Award (1R21AI123937 to O.S.A.), and CSIRO internal funding (to P.N.P.). A.B. and S.G. contributed equally to this work. Author contributions: A.B., P.N.P., and O.S.A. conceived and designed experiments; A.B., S.G., and M.L. performed all molecular and genetic experiments; I.A. performed sequencing and bioinformatic analysis; M.J.K., J.-B.D., and P.N.P. performed and analyzed data for ZIKV mosquito challenge assays; H.-H.L., H.-W.W., and C.-H.C. performed and analyzed data for in vivo mouse model ZIKV mosquito challenge assays; and A.B., S.G., M.L., I.A., H.-H.L., H.-W.W., C.-H.C., M.J.K., J.-B.D., P.N.P., and O.S.A. wrote the paper. Conflict of interest statement: A.B. and O.S.A have submitted a provisional patent application on this technology. All other authors declare no competing financial interests. This article is a PNAS Direct Submission. Data deposition: All sequencing data associated with this study are available from the National Center for Biotechnology Information Sequence Read Archive (accession no. SRP150144, BioProject accession no. PRJNA475410). The complete annotated plasmid sequence and DNA are publicly available at Addgene (plasmid no. 104968). This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1810771116/-/DCSupplemental.Attached Files
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Supplemental Material - pnas.1810771116.sapp.pdf
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Additional details
- Alternative title
- Engineered resistance to Zika virus in transgenic Ae. aegypti expressing a polycistronic cluster of synthetic miRNAs
- PMCID
- PMC6397566
- Eprint ID
- 90488
- Resolver ID
- CaltechAUTHORS:20181030-080547581
- 5K22AI113060
- NIH
- 1R21AI123937
- NIH
- Commonwealth Scientific and Industrial Research Organisation (CSIRO)
- Created
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2018-10-30Created from EPrint's datestamp field
- Updated
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2022-03-01Created from EPrint's last_modified field