School of Biological Sciences, University of Aberdeen, Aberdeen, UK.
Research Laboratories, Bambino Gesù Children's Hospital, Rome, Italy.
Sci Rep. 2020 Jul 24;10(1):12422. doi: 10.1038/s41598-020-69068-x.
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-dsRNA injected larvae, relative to the level seen in larvae injected with naked dsRNA alone. These results suggest that the use of functionalised CNTs for dsRNA delivery could increase the efficacy of RNA interference in insect pest species.
在这项研究中，评估了树枝状聚合物包覆的碳纳米管（CNTs）作为 dsRNA 递药载体在赤拟谷盗中的应用。暴露于低剂量的聚酰胺-胺树枝状大分子碳纳米管（PAMAM-CNTs）不会影响赤拟谷盗幼虫的死亡率。注射 PAMAM-CNTs 后，关键凋亡因子 Dronc（Tc12580）、Dredd（Tcn-样，Tc014026）和 Buffy（Tcinhib apop1）的表达没有改变，这些因子可以作为毒性指标。当幼虫注射与 CNTs 结合的双链 RNA（PAMAM-CNT-dsRNA）时，两个靶基因α-微管蛋白和线粒体 RNA 聚合酶（mtpol）的敲低水平与单独注射靶 dsRNA 相比显著增加。PAMAM-CNTs 在细胞空泡和细胞核中可见。与单独注射裸露 dsRNA 的幼虫相比，在注射 PAMAM-CNT-dsRNA 的幼虫中发现了一部分出现翼泡表型的增加。这些结果表明，使用功能化 CNTs 进行 dsRNA 递药可以提高 RNAi 在昆虫害虫物种中的功效。
