Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum

Catriona H Edwards, Craig Christie, Andrea Masotti, Antonella Celluzzi, Andrea Caporali, Ewan M Campbell* (Corresponding Author)

*Corresponding author for this work

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Abstract

In this study, the use of dendrimer-coated carbon nanotubes (CNTs) as a delivery vehicle for dsRNA was assessed in Tribolium castaneum. Exposure to low dosages of polyamidoamine dendrimer carbon nanotubes (PAMAM-CNTs) did not affect T. castaneum larval mortality. Expression of key apoptotic factors, Dronc (Tc12580), Dredd (Tcn-like, Tc014026) and Buffy, (Tcinhib apop1), which can act as toxicity indicators, were not altered in T. castaneum larvae following injection of PAMAM-CNTs. The level of knockdown of two target genes, α-tubulin and mitochondrial RNA polymerase (mtpol), were significantly increased when larvae were injected with double-stranded RNA bound to CNTs (PAMAM-CNT-dsRNA), compared to those injected with target dsRNA alone. PAMAM-CNTs were visualised in cellular vacuoles and in the cell nucleus. Increase occurrence of a blistered wing phenotype was found in a subset of PAMAM-CNT-dsRNAαtub injected larvae, relative to the level seen in larvae injected with naked dsRNAαtub alone. These results suggest that the use of functionalised CNTs for dsRNA delivery could increase the efficacy of RNA interference in insect pest species.

Original languageEnglish
Article number12422
Number of pages11
JournalScientific Reports
Volume10
DOIs
Publication statusPublished - 24 Jul 2020

Bibliographical note

Thanks to Dr Alan S. Bowman at the Institute for Biological and Environmental Sciences, University of Aberdeen for providing facilities and laboratory equipment for insect work and to Kevin S. Mackenzie and staff at the Microscopy and Histology Core Facility at the University of Aberdeen for TEM preparations. Scottish Crucible Project Award 2014 provided financial support for this research. CHE was supported by a Knowledge Transfer Network BBSRC Industrial Case (BB/L502467/1) studentship. CRC was supported by a KTN BBSRC CASE studentship (BB/M503526/1). AM and AC were supported by the Italian Ministry of Health (RF-PE-2011-02347026). EMC was supported by European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 613960 (SMARTBEES) and Veterinary Medicines Directorate, Department for Environment Food & Rural Affairs (Project # VM0517).

Keywords

  • animal physiology
  • nanobiotechnology
  • RNAi
  • CELLS
  • NANOMATERIALS
  • TOXICITY
  • MECHANISMS
  • AEDES-AEGYPTI
  • LARVAL
  • NANOPARTICLES
  • RNA INTERFERENCE KNOCKDOWN
  • DEGRADATION
  • ORAL RNAI

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