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 | |
| State | Published - Aug 1 2020 |
Bibliographical note
Publisher Copyright:© 2020 Wiley Periodicals, Inc.
Funding
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. 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.
| Funders | Funder number |
|---|---|
| Center for Arthropod Management Technologies | |
| Corteva Agriscience | |
| Dupont Pioneer | |
| National Science Foundation I/UCRC | |
| UCRC | |
| US Department of Agriculture | |
| National Science Foundation (NSF) | 1821936 |
| U.S. Department of Agriculture | 2353057000 |
| National Institute of Food and Agriculture | |
| Center for Arthropod Management Technologies | IIP‐1821936 |
Keywords
- Cellfectin II
- RNA interference
- Spodoptera frugiperda
- double-stranded RNA
ASJC Scopus subject areas
- Physiology
- Biochemistry
- Insect Science