Chitosan nanoparticles help double-stranded RNA escape from endosomes and improve RNA interference in the fall armyworm, Spodoptera frugiperda

Dhandapani Gurusamy, Kanakachari Mogilicherla, Subba R. Palli

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

RNA interference (RNAi) is a promising technology for the development of next-generation insect pest control products. Though RNAi is efficient and systemic in coleopteran insects, it is inefficient and variable in lepidopteron insects. In this study, we explored the possibility of improving RNAi in the fall armyworm (FAW), Spodoptera frugiperda by conjugating double-stranded RNA (dsRNA) with biodegradable chitosan (Chi). dsRNA conjugated with chitosan was protected from degradation by endonucleases present in Sf9 cell-conditioned medium, hemolymph, and midgut lumen contents collected from the FAW larvae. Chi–dsRNA complexes showed reduced accumulation in the endosomes of Sf9 cells and FAW tissues. Exposing chitosan formulated dsRNA in Sf9 cells and the tissues induced a significant knockdown of endogenous genes. Chi–dsIAP fed to FAW larvae induced knockdown of iap gene, growth retardation, and mortality. Processing of dsRNA into small interfering RNA was detected with chitosan-conjugated 32P-UTP-labeled ds green fluorescent protein in Sf9 cells and FAW larval tissues. Overall, these data suggest that dsRNA conjugated with chitosan helps dsRNA escape from the endosomes and improves RNAi efficiency in FAW cells and tissues.

Original languageEnglish
Article numbere21677
JournalArchives of Insect Biochemistry and Physiology
Volume104
Issue number4
DOIs
StatePublished - Aug 1 2020

Bibliographical note

Funding Information:
The authors thank Dr. Amit Sethi from Corteva Agriscience, DuPont–Pioneer, Dr. Guo‐Qing Tang from Syngenta, and Dr. Sven Geibel from Bayer CropScience for their kind discussions and advice. We acknowledge Mr. Jeffrey L. Howell, Senior Technician and Dr. Xien Chen, Postdoctoral Research Scholar in Palli Lab for the supply of FAW neonates and larvae for this experiment and Dr. Ramesh Kumar Dhandapani for help with DLS analysis. This material is based on the work supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies, under grant no. IIP‐1821936, and by industry partners, and the National Institute of Food and Agriculture, US Department of Agriculture (under HATCH Project 2353057000).

Funding Information:
The authors thank Dr. Amit Sethi from Corteva Agriscience, DuPont?Pioneer, Dr. Guo-Qing Tang from Syngenta, and Dr. Sven Geibel from Bayer CropScience for their kind discussions and advice. We acknowledge Mr. Jeffrey L. Howell, Senior Technician and Dr. Xien Chen, Postdoctoral Research Scholar in Palli Lab for the supply of FAW neonates and larvae for this experiment and Dr. Ramesh Kumar Dhandapani for help with DLS analysis. This material is based on the work supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies, under grant no. IIP-1821936, and by industry partners, and the National Institute of Food and Agriculture, US Department of Agriculture (under HATCH Project 2353057000).

Publisher Copyright:
© 2020 Wiley Periodicals, Inc.

Keywords

  • RNA interference
  • chitosan
  • double-stranded RNA
  • fall armyworm

ASJC Scopus subject areas

  • Physiology
  • Biochemistry
  • Insect Science

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