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伊蚊、按蚊和库蚊(双翅目:蚊科)中凋亡抑制因子转录本的表达谱及RNA干扰沉默

Expression Profiles and RNAi Silencing of Inhibitor of Apoptosis Transcripts in Aedes, Anopheles, and Culex Mosquitoes (Diptera: Culicidae).

作者信息

Puglise Jason M, Estep Alden S, Becnel James J

出版信息

J Med Entomol. 2016 Mar;53(2):304-14. doi: 10.1093/jme/tjv191.

Abstract

Effective mosquito control is vital to curtail the devastating health effects of many vectored diseases. RNA interference (RNAi)-mediated control of mosquitoes is an attractive alternative to conventional chemical pesticides. Previous studies have suggested that transcripts for inhibitors of apoptosis (IAPs) may be good RNAi targets. To revisit and extend previous reports, we examined the expression of Aedes aegypti (L.) IAPs (AaeIAPs) 1, 2, 5, 6, 9, and a viral IAP-associated factor (vIAF) as well as Anopheles quadrimaculatus Say and Culex quinquefasciatus Say IAP1 homologs (AquIAP1 and CquIAP1) in adult females. Expression profiles of IAPs suggested that some older female mosquitoes had significantly higher IAP mRNA levels when compared to the youngest ones. Minor differences in expression of AaeIAPs were observed in mosquitoes that imbibed a bloodmeal, but the majority of the time points (up to 48 h) were not significantly different. Although in vitro experiments with the Ae. aegypti Aag-2 cell line demonstrated that the various AaeIAPs could be effectively knocked down within one day after dsRNA treatment, only Aag-2 cells treated with dsIAP1 displayed apoptotic morphology. Gene silencing and mortality were also evaluated after topical application and microinjection of the same dsRNAs into female Ae. aegypti. In contrast to previous reports, topical administration of dsRNA against AaeIAP1 did not yield a significant reduction in gene expression or increased mortality. Knockdown of IAP1 and other IAPs by microinjection did not result in significant mortality. In toto, our findings suggest that IAPs may not be suitable RNAi targets for controlling adult mosquito populations.

摘要

有效的蚊虫控制对于减少许多媒介传播疾病对健康造成的毁灭性影响至关重要。RNA干扰(RNAi)介导的蚊虫控制是传统化学杀虫剂的一种有吸引力的替代方法。先前的研究表明,凋亡抑制因子(IAPs)的转录本可能是良好的RNAi靶点。为了重新审视和扩展先前的报告,我们检测了埃及伊蚊(Aedes aegypti,L.)的IAPs(AaeIAPs)1、2、5、6、9和一种病毒IAP相关因子(vIAF),以及四斑按蚊(Anopheles quadrimaculatus,Say)和致倦库蚊(Culex quinquefasciatus,Say)IAP1同源物(AquIAP1和CquIAP1)在成年雌蚊中的表达。IAPs的表达谱表明,一些较老的雌蚊与最年轻的雌蚊相比,其IAP mRNA水平显著更高。在吸食血餐的蚊虫中观察到AaeIAPs表达的微小差异,但大多数时间点(长达48小时)没有显著差异。尽管对埃及伊蚊Aag-2细胞系进行的体外实验表明,在dsRNA处理后一天内,各种AaeIAPs均可被有效敲低,但只有用dsIAP1处理的Aag-2细胞显示出凋亡形态。在用相同dsRNAs对雌性埃及伊蚊进行局部应用和显微注射后,也评估了基因沉默和死亡率。与先前的报告相反,局部应用针对AaeIAP1的dsRNA并没有导致基因表达的显著降低或死亡率的增加。通过显微注射敲低IAP1和其他IAPs并没有导致显著的死亡率。总体而言,我们的研究结果表明,IAPs可能不是控制成年蚊虫种群的合适RNAi靶点。

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