involvement of clathrin‐mediated endocytosis and two Sid‐1‐like transmembrane proteins in double‐stranded RNA uptake in the Colorado potato beetle midgut

RNA interference (RNAi) is a powerful tool in entomology and shows promise as a crop protection strategy, but variability in its efficiency across different insect species limits its applicability. For oral uptake of the double‐stranded RNA (dsRNA), the RNAi trigger, two different mechanisms are kno...

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Published in:Insect molecular biology Vol. 25; no. 3; pp. 315 - 323
Main Authors: Cappelle, K., de Oliveira, C. F. R., Van Eynde, B., Christiaens, O., Smagghe, G.
Format: Journal Article
Language:English
Published: England Published for the Royal Entomological Society by Blackwell Scientific Publications 01.06.2016
Blackwell Publishing Ltd
Subjects:
adenosinetriphosphatase
Animals
clathrin
Clathrin-Coated Vesicles - metabolism
Coleoptera - genetics
Coleoptera - metabolism
Colorado potato beetle
double-stranded RNA
double-stranded RNA uptake
Endocytosis
entomology
Gastrointestinal Tract - metabolism
genes
Genes, Insect
Genes, Reporter
hydrophobicity
Insect Proteins - metabolism
Insects
Leptinotarsa decemlineata
Membrane Transport Proteins - metabolism
midgut
phylogeny
Plant protection
Potatoes
RNA Interference
RNA, Double-Stranded - metabolism
Solanum tuberosum
systemic RNA interference deficient-1 (sid-1)
transmembrane proteins
endocytosis
RNA interference
double-stranded RNA uptake
Colorado potato beetle
systemic RNA interference deficient-1 (sid-1)
ISSN:0962-1075, 1365-2583
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Summary:RNA interference (RNAi) is a powerful tool in entomology and shows promise as a crop protection strategy, but variability in its efficiency across different insect species limits its applicability. For oral uptake of the double‐stranded RNA (dsRNA), the RNAi trigger, two different mechanisms are known: systemic RNA interference deficient‐1 (Sid‐1) transmembrane channel‐mediated uptake and clathrin‐mediated endocytosis. So far, a wide range of experiments has been conducted, confirming the involvement of one of the pathways in dsRNA uptake, but never both pathways in the same species. We investigated the role of both pathways in dsRNA uptake in the Colorado potato beetle, Leptinotarsa decemlineata, known to have an efficient RNAi response. Through RNAi‐of‐RNAi experiments, we demonstrated the contribution of two different sid‐1‐like (sil) genes, silA and silC, and clathrin heavy chain and the 16kDa subunit of the vacuolar H⁺ ATPase (vha16), elements of the endocytic pathway, to the RNAi response. Furthermore, the sid‐1‐like genes were examined through phylogenetic and hydrophobicity analysis. This article reports for the first time on the involvement of two pathways in dsRNA uptake in an insect species and stresses the importance of evaluating both pathways through a well‐devised reporter system in any future experiments on cellular dsRNA uptake.
Bibliography:http://dx.doi.org/10.1111/imb.12222
ark:/67375/WNG-PR0GNGFB-4
istex:B1AA669AC788C5E8D32E510D6B30EFC052994C67
ArticleID:IMB12222
Special Research Fund for Scientific Research Brazilian National Council for Scientific and Technological Development
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
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ISSN:0962-1075
1365-2583
DOI:10.1111/imb.12222