Arshad Fiza, Sharma Arvind, Lu Charleen, Gulia-Nuss Monika
Department of Biochemistry and Molecular Biology, University of Nevada, Reno, NV 89557, USA.
Insects. 2021 Jul 13;12(7):634. doi: 10.3390/insects12070634.
RNA-interference (RNAi) is a standard technique for functional genomics in adult mosquitoes. However, RNAi in immature, aquatic mosquito stages has been challenging. Several studies have shown successful larval RNAi, usually in combination with a carrier molecule. Except for one study in malaria mosquito, , none of the previous studies has explored RNAi in mosquito pupae. Even in the study that used RNAi in pupae, double stranded RNA (dsRNA) was introduced by microinjection. Here, we describe a successful method by soaking pupae in water containing dsRNA without any carrier or osmotic challenge. The knockdown persisted into adulthood. We expect that this simple procedure will be useful in the functional analysis of genes that highly express in pupae or newly emerged adults.
RNA干扰（RNAi）是成年蚊子功能基因组学的标准技术。然而，在未成熟的水生蚊子阶段进行RNAi一直具有挑战性。几项研究已表明幼虫RNAi成功，通常是与载体分子联合使用。除了一项针对疟蚊的研究外，之前的研究均未探索过在蚊子蛹期进行RNAi。即使在那项对蛹期使用RNAi的研究中，双链RNA（dsRNA）也是通过显微注射引入的。在此，我们描述了一种成功的方法，即将蛹浸泡在含有dsRNA的水中，无需任何载体或渗透压刺激。这种基因敲低效果持续到成年期。我们期望这一简单程序将有助于对在蛹期或新羽化的成虫中高表达的基因进行功能分析。
