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RNA Interference of the Muscle Actin Gene in Bed Bugs: Exploring Injection Versus Topical Application for dsRNA Delivery

机译:臭虫中肌动蛋白基因的RNA干扰:探索注射与局部应用dsRNA传递。

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摘要

Bed bugs are one the most troublesome household pests that feed primarily on human blood. RNA interference (RNAi) is currently being pursued as a potential tool for insect population management and has shown efficacy against some phytophagous insects. We evaluated the different techniques to deliver dsRNA specific to bed bug muscle actin (dsactin) into bed bugs. Initially, stability of dsRNA in human blood was studied to evaluate the feasibility of feeding method. Adult bed bugs were injected with dsRNA between last thoracic segment and first abdominal segment on the ventral side, with a dose of 0.2 µg dsactin per insect. In addition to injection, dsactin was mixed in acetone and treated topically in the abdomens of fifth stage nymphs. We found the quick degradation of dsRNA in blood. Injection of dsactin caused significant depletion of actin transcripts and substantial reduction in oviposition and lethality in female adults. Topically treated dsRNA in fifth stage nymphs had no effect on actin mRNA expression and survival. Our results demonstrated that injection is a reliable method of dsRNA delivery into bed bugs while topical treatment was not successful. This research provides an understanding on effective delivery methods of dsRNA into bed bugs for functional genomics research and feasibility of the RNAi based molecules for pest management purposes.
机译:臭虫是最麻烦的家庭害虫之一,主要以人类血液为食。 RNA干扰(RNAi)当前正作为一种潜在的昆虫种群管理工具而被采用,并已显示出对某些植物吞噬昆虫的功效。我们评估了将臭虫肌肉肌动蛋白(dsactin)特有的dsRNA传递到臭虫中的不同技术。最初,研究了人血液中dsRNA的稳定性,以评估喂养方法的可行性。在臭虫的最后一个胸节和腹部的第一个腹节之间,向成年臭虫注射dsRNA,每只昆虫的剂量为0.2 µg dsactin。除注射外,将双actin混合在丙酮中,并在第五阶段若虫的腹部局部治疗。我们发现血液中dsRNA迅速降解。注射双actactin会导致肌动蛋白转录本的大量消耗,以及成年女性的排卵和致死率大大降低。在第五阶段若虫中局部处理的dsRNA对肌动蛋白mRNA表达和存活没有影响。我们的结果表明,注射是将dsRNA输送到臭虫中的可靠方法,而局部治疗并不成功。这项研究提供了关于将dsRNA有效地递送到臭虫中以进行功能基因组学研究的理解,以及基于RNAi的分子用于有害生物管理的可行性。

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