Gene delivery in mosquitos with a vesicular stomatitis virus vector

Gain- and loss-of-function techniques are essential for comprehensive investigation of gene function. In mosquitoes, effective loss-of-function (LOF) methods such as RNA interference and gene knockout are available. However, convenient and practical methodologies for gain-of-function (GOF) are currently lacking. Here, we developed the vesicular stomatitis virus (VSV) vector as a GOF delivery system, which efficiently delivers target genes in mosquito Aedes aegypti without affect their fitness. Importantly, by tactically altering the insertion site of target genes within the VSV vector, we can control the relative levels of transcription. This VSV-mediated method resulted in the corresponding phenotypic changes in mosquitoes overexpressing target genes compared to controls. Additionally, we discovered that VSV could infect various orders of insects, including fruit flies (Diptera), fall armyworms (Lepidoptera), and beetles (Coleoptera), highlighting its potential as a versatile gene delivery platform across insect species.