Lipids help double-stranded RNA in endosomal escape and improve RNA interference in the fall armyworm, Spodoptera frugiperda

Dhandapani Gurusamy; Kanakachari Mogilicherla; Jayendra Nath Shukla; Subba Reddy Palli

doi:10.1002/arch.21678

Lipids help double-stranded RNA in endosomal escape and improve RNA interference in the fall armyworm, Spodoptera frugiperda

Dhandapani Gurusamy, Kanakachari Mogilicherla, Jayendra Nath Shukla, Subba Reddy Palli

Research output: Contribution to journal › Article › peer-review

37 Scopus citations

Abstract

RNA interference (RNAi) is a valuable method for understanding the gene function and holds great potential for insect pest management. While RNAi is efficient and systemic in coleopteran insects, RNAi is inefficient in lepidopteran insects. In this study, we explored the possibility of improving RNAi in the fall armyworm (FAW), Spodoptera frugiperda cells by formulating dsRNA with Cellfectin II (CFII) transfection reagent. The CFII formulated dsRNA was protected from degradation by endonucleases present in Sf9 cells conditioned medium, hemolymph and midgut lumen contents collected from the FAW larvae. Lipid formulated dsRNA also showed reduced accumulation in the endosomes of Sf9 cells and FAW tissues. Exposing Sf9 cells and tissues to CFII formulated dsRNA caused a significant knockdown of endogenous genes. CFII formulated dsIAP fed to FAW larvae induced knockdown of iap gene, growth retardation and mortality. Processing of dsRNA into siRNA was detected in Sf9 cells and Spodoptera frugiperda larvae treated with CFII conjugated ³²P-UTP labeled dsGFP. Overall, the present study concluded that delivering dsRNA formulated with CFII transfection reagent helps dsRNA escapes from the endosomal accumulation and improved RNAi efficiency in the FAW cells and tissues.

Original language	English
Article number	e21678
Journal	Archives of Insect Biochemistry and Physiology
Volume	104
Issue number	4
DOIs	https://doi.org/10.1002/arch.21678
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 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 and Dr. Ramesh Kumar Dhadapani for help with DLS analysis. This material is based on 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). The authors declare that there are no conflict of interests.

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 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 and Dr. Ramesh Kumar Dhadapani for help with DLS analysis. This material is based on 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). The authors declare that there are no conflict of interests.

Publisher Copyright:
© 2020 Wiley Periodicals, Inc.

Keywords

Cellfectin II
RNA interference
Spodoptera frugiperda
double-stranded RNA

ASJC Scopus subject areas

Physiology
Biochemistry
Insect Science

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10.1002/arch.21678

Cite this

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title = "Lipids help double-stranded RNA in endosomal escape and improve RNA interference in the fall armyworm, Spodoptera frugiperda",

abstract = "RNA interference (RNAi) is a valuable method for understanding the gene function and holds great potential for insect pest management. While RNAi is efficient and systemic in coleopteran insects, RNAi is inefficient in lepidopteran insects. In this study, we explored the possibility of improving RNAi in the fall armyworm (FAW), Spodoptera frugiperda cells by formulating dsRNA with Cellfectin II (CFII) transfection reagent. The CFII formulated dsRNA was protected from degradation by endonucleases present in Sf9 cells conditioned medium, hemolymph and midgut lumen contents collected from the FAW larvae. Lipid formulated dsRNA also showed reduced accumulation in the endosomes of Sf9 cells and FAW tissues. Exposing Sf9 cells and tissues to CFII formulated dsRNA caused a significant knockdown of endogenous genes. CFII formulated dsIAP fed to FAW larvae induced knockdown of iap gene, growth retardation and mortality. Processing of dsRNA into siRNA was detected in Sf9 cells and Spodoptera frugiperda larvae treated with CFII conjugated 32P-UTP labeled dsGFP. Overall, the present study concluded that delivering dsRNA formulated with CFII transfection reagent helps dsRNA escapes from the endosomal accumulation and improved RNAi efficiency in the FAW cells and tissues.",

keywords = "Cellfectin II, RNA interference, Spodoptera frugiperda, double-stranded RNA",

author = "Dhandapani Gurusamy and Kanakachari Mogilicherla and Shukla, {Jayendra Nath} and Palli, {Subba Reddy}",

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 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 and Dr. Ramesh Kumar Dhadapani for help with DLS analysis. This material is based on 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). The authors declare that there are no conflict of interests. 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 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 and Dr. Ramesh Kumar Dhadapani for help with DLS analysis. This material is based on 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). The authors declare that there are no conflict of interests. Publisher Copyright: {\textcopyright} 2020 Wiley Periodicals, Inc.",

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AU - Shukla, Jayendra Nath

AU - Palli, Subba Reddy

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N2 - RNA interference (RNAi) is a valuable method for understanding the gene function and holds great potential for insect pest management. While RNAi is efficient and systemic in coleopteran insects, RNAi is inefficient in lepidopteran insects. In this study, we explored the possibility of improving RNAi in the fall armyworm (FAW), Spodoptera frugiperda cells by formulating dsRNA with Cellfectin II (CFII) transfection reagent. The CFII formulated dsRNA was protected from degradation by endonucleases present in Sf9 cells conditioned medium, hemolymph and midgut lumen contents collected from the FAW larvae. Lipid formulated dsRNA also showed reduced accumulation in the endosomes of Sf9 cells and FAW tissues. Exposing Sf9 cells and tissues to CFII formulated dsRNA caused a significant knockdown of endogenous genes. CFII formulated dsIAP fed to FAW larvae induced knockdown of iap gene, growth retardation and mortality. Processing of dsRNA into siRNA was detected in Sf9 cells and Spodoptera frugiperda larvae treated with CFII conjugated 32P-UTP labeled dsGFP. Overall, the present study concluded that delivering dsRNA formulated with CFII transfection reagent helps dsRNA escapes from the endosomal accumulation and improved RNAi efficiency in the FAW cells and tissues.

AB - RNA interference (RNAi) is a valuable method for understanding the gene function and holds great potential for insect pest management. While RNAi is efficient and systemic in coleopteran insects, RNAi is inefficient in lepidopteran insects. In this study, we explored the possibility of improving RNAi in the fall armyworm (FAW), Spodoptera frugiperda cells by formulating dsRNA with Cellfectin II (CFII) transfection reagent. The CFII formulated dsRNA was protected from degradation by endonucleases present in Sf9 cells conditioned medium, hemolymph and midgut lumen contents collected from the FAW larvae. Lipid formulated dsRNA also showed reduced accumulation in the endosomes of Sf9 cells and FAW tissues. Exposing Sf9 cells and tissues to CFII formulated dsRNA caused a significant knockdown of endogenous genes. CFII formulated dsIAP fed to FAW larvae induced knockdown of iap gene, growth retardation and mortality. Processing of dsRNA into siRNA was detected in Sf9 cells and Spodoptera frugiperda larvae treated with CFII conjugated 32P-UTP labeled dsGFP. Overall, the present study concluded that delivering dsRNA formulated with CFII transfection reagent helps dsRNA escapes from the endosomal accumulation and improved RNAi efficiency in the FAW cells and tissues.

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KW - double-stranded RNA

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Lipids help double-stranded RNA in endosomal escape and improve RNA interference in the fall armyworm, Spodoptera frugiperda

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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