Rehydration Driven RNAi: A Novel Approach for Effectively Delivering dsRNA to Mosquito Larvae
Rehydration Driven RNAi: A Novel Approach for Effectively Delivering dsRNA to Mosquito Larvae
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DOI：
10.1603/me11122
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发表时间：
2012-01-01
影响因子：
2.1
通讯作者：
Denlinger, David L.
中科院分区：
文献类型：
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作者：
Lopez-Martinez, Giancarlo;Meuti, Megan;Denlinger, David L.
蚊子幼虫柔软的身体和水生生境为应用标准的RNA干扰技术沉默靶基因的表达提出了挑战。在这里，我们描述了一种新的技术，通过利用蚊子幼虫的脱水耐受性将双链RNA传递到蚊子幼虫体内。幼虫在氯化钠溶液中脱水，然后在含有双链RNA的水中复水。利用淡色库蚊幼虫进行诱变试验。我们通过下调编码热休克蛋白90的基因的表达来证明这一原理。这种基因敲除一直持续到蛹阶段和成虫，在成年生命的第三天，大约77%的基因敲除仍然明显。我们预计，这一相对简单的程序将被证明对抑制这种蚊子幼虫和其他蚊子中其他基因的表达也是有用的。
The soft bodies and aquatic habitats of mosquito larvae pose a challenge for applying standard RNA-interference techniques to silence expression of target genes. Here we describe a novel technique for delivering double-stranded RNA into mosquito larvae by exploiting the larva's dehydration tolerance. Larvae were dehydrated in a NaCl solution and then rehydrated in water containing double-stranded RNA. Using larvae of Culex pipiens (L.) we demonstrated the principle by knocking down expression of the gene encoding heat shock protein 90. The knockdown persisted through the pupal stage and into adulthood, with a knockdown of approximate to 77% still evident on the third day of adult life. We anticipate that this relatively simple procedure will prove useful for knocking down expression of other genes as well, in larvae of this mosquito and in others.