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

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Tytuł:: Lipids help double-stranded RNA in endosomal escape and improve RNA interference in the fall armyworm, Spodoptera frugiperda.
Autorzy:: Gurusamy D; Department of Entomology, University of Kentucky, Lexington, Kentucky, 40546, USA.
Mogilicherla K; Department of Entomology, University of Kentucky, Lexington, Kentucky, 40546, USA.
Shukla JN; Department of Entomology, University of Kentucky, Lexington, Kentucky, 40546, USA.
Palli SR; Department of Entomology, University of Kentucky, Lexington, Kentucky, 40546, USA.
Źródło:: Archives of insect biochemistry and physiology [Arch Insect Biochem Physiol] 2020 Aug; Vol. 104 (4), pp. e21678. Date of Electronic Publication: 2020 Apr 15.
Typ publikacji:: Journal Article
Język:: English
Imprint Name(s):: Publication: New York, NY : Wiley
Original Publication: New York : Alan R. Liss, c1983-
MeSH Terms:: RNA Interference*
Liposomes/*administration & dosage
RNA, Double-Stranded/*administration & dosage
Spodoptera/*drug effects
Animals ; Endosomes ; Insect Control/methods ; Larva/drug effects ; RNA, Small Interfering ; Sf9 Cells ; Spodoptera/growth & development ; Transfection/methods
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Grant Information:: IIP-1821936 National Science Foundation; 2353057000 USDA
Contributed Indexing:: Keywords: Cellfectin II; RNA interference; Spodoptera frugiperda; double-stranded RNA
Substance Nomenclature:: 0 (Liposomes)
0 (RNA, Double-Stranded)
0 (RNA, Small Interfering)
Entry Date(s):: Date Created: 20200417 Date Completed: 20200929 Latest Revision: 20200929
Update Code:: 20240104
DOI:: 10.1002/arch.21678
PMID:: 32297364
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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 32 P-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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