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

doi:10.1002/arch.21677

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 journal › Article › peer-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 language	English
Article number	e21677
Journal	Archives of Insect Biochemistry and Physiology
Volume	104
Issue number	4
DOIs	https://doi.org/10.1002/arch.21677
State	Published - 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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10.1002/arch.21677

Cite this

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

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.",

keywords = "RNA interference, chitosan, double-stranded RNA, fall armyworm",

author = "Dhandapani Gurusamy and Kanakachari Mogilicherla and Palli, {Subba R.}",

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: {\textcopyright} 2020 Wiley Periodicals, Inc.",

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AU - Palli, Subba R.

N1 - 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.

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N2 - 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.

AB - 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.

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

Abstract

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Keywords

ASJC Scopus subject areas

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