中华按蚊RNAi技术平台的建立及应用

Establishment and Application of Anopheles sinensis RNAi Technology Platform

【目的】建立一套经济、便捷及高效的RNAi技术平台。【方法】以中华按蚊(Anopheles sinensis)为模式物种,从材料固定、注射部位、注射剂量等方面进行改进,并利用该平台沉默中华按蚊黑色素代谢途径中在蛹期表达的一个关键基因,以验证该技术平台的操作性。【结果】建立了中华按蚊显微注射平台,通过摸索和改进注射操作技术,利用该平台成功地实现了幼虫、蛹和成蚊的显微注射;同时,有效地沉默了该关键基因的表达,获得了黑化缺陷的蛹表型。【结论】该平台体系的建立,不仅为中华按蚊功能基因的研究提供了分析手段,同时也为其他涉及显微注射的技术体系提供了参考。

[Purposes]The paper aims to establish an economical,convenient and efficient RNAi technology platform.[Methods]Anopheles sinensis was used as the model to improve the RNAi operation,including mosquito fixation,injection site and dose,etc.And the expression of a key gene in An.sinensis melanin metabolic pathway in the pupal stage was silenced via the platform to verify the operability of the platform.[Findings]The platform for A.sinensis microinjection was established.After the improvement of the injection operational technology,larva was injected successfully,pupa and adult and efficiently silenced the expression of the key gene via the platform,and finally the non-melanic pupae were obtained.[Conclusions]The establishment of the platform system not only provided us an analytical tool for the study of A.sinensis gene function,but also provided a reference for other technology systems involved in microinjection.