Systemic gene knockdown in Camponotus floridanus workers by feeding of dsRNA
Systemic gene knockdown in Camponotus floridanus workers by feeding of dsRNA
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DOI：
10.1007/s00040-013-0314-6
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发表时间：
2013-08
期刊：
影响因子：
1.3
通讯作者：
C. Ratzka;R. Gross;H. Feldhaar
中科院分区：
文献类型：
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作者：
C. Ratzka;R. Gross;H. Feldhaar
RNA干扰（RNAi）技术使得研究社会昆虫中的特定基因功能成为可能，否则很难进行遗传操作。最近对七种蚂蚁的基因组进行了测序，使这些蚁科成员可用于击倒研究。然而，为此目的，RNAi技术首先需要适应于在蚂蚁中的应用。对其他昆虫的研究表明，RNAi的有效性具有很强的物种特异性，并且可能取决于几个实验参数，例如昆虫的研究阶段，靶基因和/或dsRNA递送方法。通过饲喂dsRNA在蚂蚁中进行RNAi是优选的方法，因为与注射dsRNA相比，可以在个体中实现敲减而不干扰动物的生理学。在这里，我们提出了一个协议，基因敲除蚁科喂养的dsRNA工人的动物。在佛罗里达弓背蚁（Camponotusfloridanus）工蚁体内，肽聚糖识别蛋白基因PGRP-LB的表达被有效地敲低。此外，我们描述了一种相对便宜的方法来提取dsRNA的细菌，以获得大量的喂养实验所需的。
RNA interference (RNAi) technology enables to study specific gene functions also in social insects, which are otherwise difficult to access for genetic manipulations. The recent sequencing of the genomes from seven ant species made these members of the Formicidae available for knockdown studies. However, for this purpose the RNAi technology first needs to be adapted for application in ants. Studies on other insects show that the effectiveness of RNAi is quite species-specific and can depend on several experimental parameters such as the investigated stage of the insect, the target gene and/or the dsRNA delivery method. RNAi in ants through feeding of dsRNA is a preferable approach, since knockdown can be achieved in individuals without interfering with the animal’s physiology in contrast to injection of dsRNA. Here, we present a protocol for gene knockdown in Formicidae by feeding of dsRNA to worker animals. The expression of apeptidoglycan recognition proteingene,PGRP-LB, was efficiently knocked down in the body ofCamponotusfloridanusworker ants. Moreover, we describe a relatively cheap method to extract dsRNA from bacteria in order to obtain large quantities needed for feeding experiments.