Baculovirus as an efficient vector for gene delivery into mosquitoes

• Published in: Scientific reports 2018-12, Vol.8 (1), p.17778-14, Article 17778
• Main Authors: Naik, Nenavath Gopal, Lo, Yu-Wen, Wu, Tzong-Yuan, Lin, Chang-Chi, Kuo, Szu-Cheng, Chao, Yu-Chan
• Format: Article
• Language: English
• Subjects: 42; 42/44; 631/1647/2300/1850; 631/326/2522; Animals; Baculoviridae - genetics; Cell Line; Cell lines; Chlorocebus aethiops; Culicidae; Culicidae - genetics; Culicidae - virology; Deoxyribonucleic acid; Disease Vectors; DNA; Exo-a-sialidase; Gene expression; Gene Expression - genetics; Gene regulation; Gene transfer; Gene Transfer Techniques; HEK293 Cells; Humanities and Social Sciences; Humans; Influenza; Larva - genetics; Larva - virology; Mosquito Vectors - genetics; Mosquitoes; multidisciplinary; Neuraminidase - genetics; Nucleopolyhedroviruses - genetics; Orthomyxoviridae - genetics; Science; Science (multidisciplinary); Transfection; Transfection - methods; Vero Cells
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-018-35463-8

Efficient gene delivery technologies play an essential role in the gene functional analyses that are necessary for basic and applied researches. Mosquitoes are ubiquitous insects, responsible for transmitting many deadly arboviruses causing millions of human deaths every year. The lack of efficient and flexible gene delivery strategies in mosquitoes are among the major hurdles for the study of mosquito biology and mosquito-pathogen interactions. We found that Autographa californica multiple nucleopolyhedrovirus (AcMNPV), the type baculovirus species, can efficiently transduce mosquito cells without viral propagation, allowing high level gene expression upon inducement by suitable promoters without obvious negative effects on cell propagation and viability. AcMNPV transduces into several mosquito cell types, efficiently than in commonly used mammalian cell lines and classical plasmid DNA transfection approaches. We demonstrated the application of this system by expressing influenza virus neuraminidase (NA) into mosquito hosts. Moreover, AcMNPV can transduce both larvae and adults of essentially all blood-sucking mosquito genera, resulting in bright fluorescence in insect bodies with little or no tissue barriers. Our experiments establish baculovirus as a convenient and powerful gene delivery vector in vitro and in vivo that will greatly benefit research into mosquito gene regulation, development and the study of mosquito-borne viruses.