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口服 dsRNA 可诱导亚洲长角牛,Anoplophora glabripennis 靶基因的敲低和死亡。

Orally delivered dsRNA induces knockdown of target genes and mortality in the Asian long-horned beetle, Anoplophora glabripennis.

机构信息

Department of Entomology, College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky.

USDA ARS Beneficial Insects Introduction Research Unit, Newark, Delaware.

出版信息

Arch Insect Biochem Physiol. 2020 Aug;104(4):e21679. doi: 10.1002/arch.21679. Epub 2020 Apr 15.

Abstract

The Asian long-horned beetle (ALB) Anoplophora glabripennis is a serious invasive forest pest in several countries, including the United States. Methods available to manage or eradicate this pest are extremely limited, but RNA interference (RNAi) technology is a potentially effective method to control ALB. In this study, we used sucrose feeding bioassay for oral delivery of double-strand RNA (dsRNA) to ALB larvae. P-labeled dsRNA orally delivered to ALB larvae using the sucrose droplet feeding method was processed to small interfering RNA. Feeding neonate larvae with dsRNA targeting genes coding for the inhibitor of apoptosis (IAP), vacuolar sorting protein SNF7 (SNF7), and snakeskin (SSK) induced knockdown of target genes and mortality. Feeding 2 µg of dsRNA per day for 3 days did not induce a significant decrease in the expression of target genes or mortality. However, feeding 5 or 10 µg of dsRNA per day for 3 days induced a significant decrease in the expression of target genes and 50-90% mortality. Interestingly, feeding 2.5 µg each of dsIAP plus dsSNF7, dsIAP plus dsSSK, or dsSNF7 plus dsSSK per day for 3 days induced a significant decrease in the expression of both target genes and approximately 80% mortality. Our findings demonstrate that orally delivered dsRNA induces target gene knockdown and mortality in ALB neonate larvae and RNAi technology may have the potential for effective ALB control.

摘要

亚洲长角天牛(ALB)Anoplophora glabripennis 是包括美国在内的几个国家的严重入侵森林害虫。目前可用于管理或根除这种害虫的方法极为有限,但 RNA 干扰(RNAi)技术是一种控制 ALB 的潜在有效方法。在这项研究中,我们使用蔗糖喂食生物测定法通过口服传递双链 RNA(dsRNA)来处理 ALB 幼虫。 使用蔗糖液滴喂食方法将 P 标记的 dsRNA 口服传递给 ALB 幼虫,然后将其加工成小干扰 RNA。用靶向编码凋亡抑制剂(IAP)、液泡分选蛋白 SNF7(SNF7)和蛇皮(SSK)的基因的 dsRNA 喂养幼虫,会导致靶基因的敲低和死亡率。每天喂食 2μg dsRNA 连续 3 天不会导致靶基因表达显著降低或死亡率增加。然而,每天喂食 5 或 10μg dsRNA 连续 3 天会导致靶基因表达显著降低和 50-90%的死亡率。有趣的是,每天喂食 2.5μg 每种 dsIAP 加 dsSNF7、dsIAP 加 dsSSK 或 dsSNF7 加 dsSSK 连续 3 天会导致靶基因的表达显著降低和大约 80%的死亡率。我们的研究结果表明,口服传递的 dsRNA 可诱导 ALB 幼虫的靶基因敲低和死亡率,并且 RNAi 技术可能具有有效控制 ALB 的潜力。

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