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首页> 外文期刊>International Journal for Parasitology >RNA interference in the cat flea, Ctenocephalides felis: Approaches for sustained gene knockdown and evidence of involvement of Dicer-2 and Argonaute2
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RNA interference in the cat flea, Ctenocephalides felis: Approaches for sustained gene knockdown and evidence of involvement of Dicer-2 and Argonaute2

机译：猫跳蚤的RNA干扰，Ctenocephalides Felis：持续基因敲低的方法和Dicer-2和Argonaute2的参与证据

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摘要

Effective RNA interference (RNAi) methods have been developed in many pest species, enabling exploration of gene function. Until now RNAi had not been attempted in the cat flea, Ctenocephalides felis, although the development of RNAi approaches would open up potential avenues for control of this important pest. This study aimed to establish if an RNAi response occurs in adult C. felis upon exposure to double-stranded RNA (dsRNA), which administration methods for dsRNA delivery could bring about effective gene knockdown and to investigate dynamics of any RNAi response. Knockdown of 80% of GST sigma was achieved by intrahaemoceolic microinjection of dsGST sigma but this invasive technique was associated with relatively high mortality rates. Immersing C. felis in dsGST sigma or dsDicer-2 overnight resulted in 65% knockdown of GST sigma or 60% of Dicer-2, respectively, and the degree of knockdown was not improved by increasing the dsRNA concentration in the bathing solution. Unexpectedly, the greatest degree of knockdown was achieved with the continuous administration of dsRNA in whole blood via a membrane feeding system, resulting in 96% knockdown of GST sigma within 2 days and sustained up to, at least, 7 days. Thus, unlike in many other species, the gut nucleases do not impair the RNAi response to ingested dsRNA in C. felis. A modest, but significant, upregulation of Dicer-2 and Argonaute2 was detectable 3 h after exposure to exogenous dsRNA, implicating the short-interfering RNA pathway. To our knowledge this study represents the first demonstration of experimentally induced RNAi in the cat flea as well as giving insight into how the gene knockdown response progresses. (C) 2018 The Author(s). Published by Elsevier Ltd on behalf of Australian Society for Parasitology.
机译：有效的RNA干扰（RNAi）方法是在许多害虫物种中开发的，从而能够探索基因功能。到目前为止，RNAI尚未尝试在猫跳蚤中，虽然RNAi方法的发展将开辟潜在的途径，以控制这一重要害虫。本研究旨在建立RNAi反应，如果在接触双链RNA（DSRNA）时，FELIS在暴露于双链RNA（DSRNA）中，这对于DSRNA递送的给药方法可以带来有效的基因敲低，并研究任何RNAi反应的动态。通过DSGST Sigma的肠道微调显微注射，实现了80％GST Sigma的敲低，但这种侵入性技术与相对较高的死亡率有关。浸入DSGST Sigma或DSDICER-2中的FELIS过夜导致GST Sigma的65％敲低，分别通过增加沐浴溶液中的DSRNA浓度来改善敲低程度。出乎意料地，通过膜饲养系统连续施用DSRNA在全血中连续施用最大程度的敲低，从而在2天内得到96％的GST Sigma敲低，至少持续到7天。因此，与许多其他物种不同，肠道核酸酶不会损害RNAi反应在C.Felis中摄入的DsRNA。在暴露于外源性DSRNA之后，Dicer-2和Argonaute2对Dicer-2和Argonaute2的较大，令人显着的，暗示的是，意味着短暂干扰的RNA途径。据我们所知，这项研究代表了在猫跳蚤中实验诱导的RNAi的第一次演示，并介绍了基因敲低响应的进展情况。 （c）2018提交人。由elsevier有限公司发布代表澳大利亚寄生虫学会。

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