靶向gat基因的shRNA重组微藻防控白纹伊蚊

Recombinant expression of shRNA targeting gat in microalgae for controlling Aedes albopictus

摘要:

目的 探讨利用转基因微藻防控传播登革热等疾病的伊蚊的技术可行性。方法 以伊蚊神经递质转运体γ-氨基丁酸受体基因(gamma-aminobutyric acid receptor gene, gat)为靶点，构建shRNA微藻(Microalgae)表达载体，通过电击法将其转入莱茵衣藻(Chlamydomonas reinhardtii) CC124和小球藻(Chlorella vulgaris) HOC5中，获得重组微藻，并用其饲喂伊蚊幼虫和成蚊。结果 gat-shRNA转基因藻株可显著延缓伊蚊幼虫的发育、对伊蚊幼虫和成蚊表现出明显的致死效果，致死率分别为75.56%和58.67%。伊蚊幼虫体内目标基因gat的表达量显著降低。结论 在封闭水域中投放gat-shRNA转基因微藻以抑制水域附近的蚊子种群是可行的。该策略为使用生物方法控制伊蚊，以及阻断登革热、寨卡病毒病等严重传染病的传播提供了一种新的思路。

关键词:莱茵衣藻;小球藻;登革热;伊蚊;gat基因

Abstract:

Objective To explore the technical feasibility of using transgenic microalgae to control Aedes that transmit diseases such as dengue fever.Methods Taking the neurotransmitter transporter gamma-aminobutyric acid receptor gene (gat) of Aedes as a target, we constructed a shRNA-expressing vector and then transferred the vector into Chlamydomonas reinhardtiii CC124 and Chlorella vulgaris HOC5 through electroporation. The recombinant microalgae were used to feed the larvae and adults of Aedes albopictus.Results The transgenic algae delayed the larval development and had obvious lethal effects on the larvae and adults of A. albopictus, causing the mortality rates of 75.56% and 58.67%, respectively. The expression level of gat in the larvae was significantly down-regulated.Conclusion Deploying gat-shRNA transgenic microalgae in enclosed water environments to suppress mosquito populations in the vicinity is technically feasible. This strategy provides a new perspective on using biological methods to control mosquitoes and block the transmission of severe infectious diseases such as dengue fever and Zika virus disease.

Key words:Chlamydomonas reinhardtiii;Chlorella vulgaris;dengue fever;Aedes;gat