Cited by 473Insect Resistance and GeneticsRNA Interference and Gene DeliveryInsect Pest Control Strategies

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Characterizing the Mechanism of Action of Double-Stranded RNA Activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte)

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Bolognesi, Renata, Ramaseshadri, Parthasarathy, Anderson, Jerry, Bachman, Pamela, Clinton, William, Flannagan, Ronald, Ilagan, Oliver, Lawrence, Christina, Levine, Steven, Moar, William, Mueller, Geoffrey, Tan, Jianguo, Uffman, Joshua, Wiggins, Elizabeth, Heck, Gregory, Segers, Gerrit·2012·PLoS ONE

DOI 10.1371/journal.pone.0047534PMC PMC3469495

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Abstract

RNA interference (RNAi) has previously been shown to be effective in western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) larvae via oral delivery of synthetic double-stranded RNA (dsRNA) in an artificial diet bioassay, as well as by ingestion of transgenic corn plant tissues engineered to express dsRNA. Although the RNAi machinery components appear to be conserved in Coleopteran insects, the key steps in this process have not been reported for WCR. Here we characterized the sequence of events that result in mortality after ingestion of a dsRNA designed against WCR larvae. We selected the Snf7 ortholog (DvSnf7) as the target mRNA, which encodes an essential protein involved in intracellular trafficking. Our results showed that dsRNAs greater than or equal to approximately 60 base-pairs (bp) are required for biological activity in artificial diet bioassays. Additionally, 240 bp dsRNAs containing a single 21 bp match to the target sequence were also efficacious, whereas 21 bp short interfering (si) RNAs matching the target sequence were not. This result was further investigated in WCR midgut tissues: uptake of 240 bp dsRNA was evident in WCR midgut cells while a 21 bp siRNA was not, supporting the size-activity relationship established in diet bioassays. DvSnf7 suppression was observed in a time-dependent manner with suppression at the mRNA level preceding suppression at the protein level when a 240 bp dsRNA was fed to WCR larvae. DvSnf7 suppression was shown to spread to tissues beyond the midgut within 24 h after dsRNA ingestion. These events (dsRNA uptake, target mRNA and protein suppression, systemic spreading, growth inhibition and eventual mortality) comprise the overall mechanism of action by which DvSnf7 dsRNA affects WCR via oral delivery and provides insights as to how targeted dsRNAs in general are active against insects.

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[23]Bucher G, Scholten J, Klinger M (2002) Parental RNAi in Tribolium (Coleoptera) Current Biology. 12: R85– R86. 11839285
[41]Ramaseshadri P, Segers G, R F, Clinton W, Oliver I, et al.. (2012) Suppression of a component of the ESCRT-III pathway (Snf7) by oral RNAi disrupts endosomal-autophagic pathway in the western corn rootworm (Diabrotica virgifera virgifera) larvae Plos One. Under review.

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Authors: Bolognesi, R., Ramaseshadri, P., Anderson, J., Bachman, P., Clinton, W., Flannagan, R., Ilagan, O., Lawrence, C., Levine, S., Moar, W., Mueller, G., Tan, J., Uffman, J., Wiggins, E., Heck, G., & Segers, G.
Affiliations: Monsanto (United States) (US)
Corresponding author: Ramaseshadri, Parthasarathy
ORCID: Bachman, Pamela
Flannagan, Ronald
Levine, Steven
Moar, William
Mueller, Geoffrey
Venue: PLoS ONE
Publisher: Public Library of Science (PLoS)
Published: 2012-10-11
Volume: 7
Issue: 10
Pages: e47534
ISSN: 1932-6203 (ISSN-L)
DOI: 10.1371/journal.pone.0047534
PMC: PMC3469495
OpenAlex: W2021552468
Cited by: 473
References: 42
Citations by year: 2026: 24 · 2025: 49 · 2024: 29 · 2023: 31 · 2022: 52 · 2021: 44 · 2020: 43 · 2019: 50 · 2018: 39 · 2017: 28 · 2016: 33 · 2015: 24 · 2014: 14 · 2013: 13
FWCI: 6.52
Type: journal article
Language: en
Open access: gold
doi.org
Licence: cc-by
creativecommons.org/licenses/by/4.0/ (unspecified)
Locations: PLoS ONE (publishedVersion, cc-by)
PubMed Central (publishedVersion, cc-by)
PubMed (publishedVersion)
DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Figshare (submittedVersion, cc-by)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Insect Resistance and Genetics
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Life Sciences › Agricultural and Biological Sciences › Plant Science › Insect Pest Control Strategies
Keywords: western corn rootworm, RNA interference, double-stranded RNA, siRNA, oral dsRNA delivery, systemic RNAi, insect mortality, transgenic maize, insect pest control
MeSH terms: Analysis of Variance, Animals, Base Sequence, Coleoptera, Biological Assay, Enzyme-Linked Immunosorbent Assay, Insect Control, Larva, Lethal Dose 50, Molecular Sequence Data, RNA, Double-Stranded, Blotting, Western, Reverse Transcriptase Polymerase Chain Reaction, RNA Interference, Gastrointestinal Tract, Endosomal Sorting Complexes Required for Transport, Real-Time Polymerase Chain Reaction

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Characterizing the Mechanism of Action of Double-Stranded RNA Activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte)

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