Engineered Flock House Virus for Targeted Gene Suppression Through RNAi in Fruit Flies (Drosophila melanogaster ) in Vitro and in Vivo

Frontiers in Physiology
Clauvis N T TaningGuy Smagghe

Abstract

RNA interference (RNAi) is a powerful tool to study functional genomics in insects and the potential of using RNAi to suppress crop pests has made outstanding progress. However, the delivery of dsRNA is a challenging step in the development of RNAi bioassays. In this study, we investigated the ability of engineered Flock House virus (FHV) to induce targeted gene suppression through RNAi under in vitro and in vivo condition. As proxy for fruit flies of agricultural importance, we worked with S2 cells as derived from Drosophila melanogaster embryos, and with adult stages of D. melanogaster. We found that the expression level for all of the targeted genes were reduced by more than 70% in both the in vitro and in vivo bioassays. Furthermore, the cell viability and median survival time bioassays demonstrated that the recombinant FHV expressing target gene sequences caused a significantly higher mortality (60-73% and 100%) than the wild type virus (24 and 71%), in both S2 cells and adult insects, respectively. This is the first report showing that a single stranded RNA insect virus such as FHV, can be engineered as an effective in vitro and in vivo RNAi delivery system. Since FHV infects many insect species, the described method coul...Continue Reading

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Citations

Nov 22, 2019·Neotropical Entomology·N P DiasM J Zotti
Jul 23, 2020·Pest Management Science·Rohit SharmaGuy Smagghe
Nov 12, 2019·Frontiers in Plant Science·Deise CagliariMoisés João Zotti
Aug 1, 2020·Frontiers in Plant Science·Anna KolliopoulouLuc Swevers
Oct 15, 2019·Annual Review of Entomology·Kun Yan Zhu, Subba Reddy Palli
May 7, 2020·Frontiers in Plant Science·Olivier ChristiaensGuy Smagghe
Jan 9, 2021·Annual Review of Entomology·Bryony C Bonning, Maria-Carla Saleh
Feb 2, 2021·Saudi Journal of Biological Sciences·Sayed Sartaj SohrabEsam Ibraheem Azhar
Jan 26, 2021·Journal of Infection and Public Health·Sayed Sartaj SohrabEsam Ibraheem Azhar
May 6, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sherif Aly El-KafrawyEsam Ibraheem Azhar

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Methods Mentioned

BETA
transfection
restriction digest
Fluorescence Microscopy
electrophoresis
PCR
light microscopy

Software Mentioned

BLAST
GraphPad Prism
qBase
CFX manager
NIS Elements BR

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