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

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...

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Main Authors: Catriona H. Edwards, Craig R. Christie, Andrea Masotti, Antonella Celluzzi, Andrea Caporali, Ewan M. Campbell
Format: Article
Language:English
Published: Nature Publishing Group 2020-07-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-020-69068-x
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spelling doaj-3e64dda316b246c1b89de27d9c2b11a82021-07-25T11:22:01ZengNature Publishing GroupScientific Reports2045-23222020-07-0110111110.1038/s41598-020-69068-xDendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneumCatriona H. Edwards0Craig R. Christie1Andrea Masotti2Antonella Celluzzi3Andrea Caporali4Ewan M. Campbell5School of Biological Sciences, University of AberdeenSchool of Biological Sciences, University of AberdeenResearch Laboratories, Bambino Gesù Children’s HospitalResearch Laboratories, Bambino Gesù Children’s HospitalCentre for Cardiovascular Science, University of EdinburghCentre for Genome Enabled Biology and Medicine, University of AberdeenAbstract 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.https://doi.org/10.1038/s41598-020-69068-x
collection DOAJ
language English
format Article
sources DOAJ
author Catriona H. Edwards
Craig R. Christie
Andrea Masotti
Antonella Celluzzi
Andrea Caporali
Ewan M. Campbell
spellingShingle Catriona H. Edwards
Craig R. Christie
Andrea Masotti
Antonella Celluzzi
Andrea Caporali
Ewan M. Campbell
Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum
Scientific Reports
author_facet Catriona H. Edwards
Craig R. Christie
Andrea Masotti
Antonella Celluzzi
Andrea Caporali
Ewan M. Campbell
author_sort Catriona H. Edwards
title Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum
title_short Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum
title_full Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum
title_fullStr Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum
title_full_unstemmed Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum
title_sort dendrimer-coated carbon nanotubes deliver dsrna and increase the efficacy of gene knockdown in the red flour beetle tribolium castaneum
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2020-07-01
description 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.
url https://doi.org/10.1038/s41598-020-69068-x
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