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一种新型 miRNA,miR-13664,通过靶向 CpCYP314A1 来调节淡色库蚊对溴氰菊酯的抗性。

A novel miRNA, miR-13664, targets CpCYP314A1 to regulate deltamethrin resistance in Culex pipiens pallens.

机构信息

Department of Pathogen Biology,Nanjing Medical University,Nanjing,Jiangsu 211166,PR China.

Department of Pathogen Biology,Hebei Medical University,Shijiazhuang 050017,China.

出版信息

Parasitology. 2019 Feb;146(2):197-205. doi: 10.1017/S0031182018001002. Epub 2018 Jul 3.

Abstract

Extensive insecticide use has led to the resistance of mosquitoes to these insecticides, posing a major barrier to mosquito control. Previous Solexa high-throughput sequencing of Culex pipiens pallens in the laboratory has revealed that the abundance of a novel microRNA (miRNA), miR-13664, was higher in a deltamethrin-sensitive (DS) strain than a deltamethrin-resistant (DR) strain. Real-time quantitative PCR revealed that the miR-13664 transcript level was lower in the DR strain than in the DS strain. MiR-13664 oversupply in the DR strain increased the susceptibility of these mosquitoes to deltamethrin, whereas inhibition of miR-13664 made the DS strain more resistant to deltamethrin. Results of bioinformatic analysis, quantitative reverse-transcriptase polymerase chain reaction, luciferase assay and miR mimic/inhibitor microinjection revealed CpCYP314A1 to be a target of miR-13664. In addition, downregulation of CpCYP314A1 expression in the DR strain reduced the resistance of mosquitoes to deltamethrin. Taken together, our results indicate that miR-13664 could regulate deltamethrin resistance by interacting with CpCYP314A1, providing new insights into mosquito resistance mechanisms.

摘要

大量使用杀虫剂导致蚊子对这些杀虫剂产生了抗性,这对蚊子的控制构成了重大障碍。之前在实验室对淡色库蚊进行的 Solexa 高通量测序表明,一种新型 microRNA(miRNA),miR-13664,在对溴氰菊酯敏感(DS)的品系中比在对溴氰菊酯抗性(DR)的品系中更为丰富。实时定量 PCR 显示,DR 品系中的 miR-13664 转录本水平低于 DS 品系。DR 品系中 miR-13664 的过度供应增加了这些蚊子对溴氰菊酯的敏感性,而 miR-13664 的抑制作用使 DS 品系对溴氰菊酯更具抗性。生物信息学分析、定量逆转录聚合酶链反应、荧光素酶测定和 miR 模拟物/抑制剂微注射的结果表明 CpCYP314A1 是 miR-13664 的靶标。此外,下调 DR 品系中 CpCYP314A1 的表达降低了蚊子对溴氰菊酯的抗性。综上所述,我们的研究结果表明,miR-13664 可以通过与 CpCYP314A1 相互作用来调节溴氰菊酯的抗性,为蚊子的抗性机制提供了新的见解。

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