Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests

Frontiers in Physiology
François-Xavier GilletMaria Fátima Grossi-de-Sá

Abstract

Genetically modified (GM) crops producing double-stranded RNAs (dsRNAs) are being investigated largely as an RNA interference (RNAi)-based resistance strategy against crop insect pests. However, limitations of this strategy include the sensitivity of dsRNA to insect gut nucleases and its poor insect cell membrane penetration. Working with the insect pest cotton boll weevil (Anthonomus grandis), we showed that the chimeric protein PTD-DRBD (peptide transduction domain-dsRNA binding domain) combined with dsRNA forms a ribonucleoprotein particle (RNP) that improves the effectiveness of the RNAi mechanism in the insect. The RNP slows down nuclease activity, probably by masking the dsRNA. Furthermore, PTD-mediated internalization in insect gut cells is achieved within minutes after plasma membrane contact, limiting the exposure time of the RNPs to gut nucleases. Therefore, the RNP provides an approximately 2-fold increase in the efficiency of insect gene silencing upon oral delivery when compared to naked dsRNA. Taken together, these data demonstrate the role of engineered RNPs in improving dsRNA stability and cellular entry, representing a path toward the design of enhanced RNAi strategies in GM plants against crop insect pests.

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Citations

Nov 22, 2019·Neotropical Entomology·N P DiasM J Zotti
Jun 21, 2020·Molecular Biology Reports·Mohsin AliJoy O Nwogwugwu
Jun 6, 2020·Frontiers in Plant Science·Marcos Fernando BassoMaria Fatima Grossi-de-Sa
Jun 26, 2020·Frontiers in Plant Science·Jörg Romeis, Franco Widmer
Dec 21, 2017·PloS One·Rayssa Almeida GarciaMaria Fatima Grossi-de-Sa
Jan 29, 2019·Frontiers in Physiology·Elise VogelJozef Vanden Broeck
May 7, 2020·Frontiers in Plant Science·Olivier ChristiaensGuy Smagghe
Jan 5, 2020·Nature Communications·Shao-Ru WangXiang Zhou
Oct 15, 2019·Annual Review of Entomology·Kun Yan Zhu, Subba Reddy Palli
Oct 8, 2020·Neotropical Entomology·Ricardo SalvadorH Esteban Hopp
Oct 26, 2020·Pest Management Science·Ritesh G JainNeena Mitter
Aug 25, 2020·Chembiochem : a European Journal of Chemical Biology·Wes BrownAlexander Deiters
Jan 14, 2021·Pest Management Science·Kristopher SilverKun Yan Zhu
Jun 10, 2020·Current Opinion in Insect Science·Rachel M Wiltshire, Molly Duman-Scheel

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Methods Mentioned

BETA
electrophoretic mobility shift assay
affinity purification
electrophoresis
cleavage assay
confocal microscopy
dissection
PCR

Software Mentioned

Fiji
qBasePlus
time PCR Miner
Real
ImageJ

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