A modified feeding RNAi method for simultaneous knock-down of more than one gene in Caenorhabditis elegans
A modified feeding RNAi method for simultaneous knock-down of more than one gene in Caenorhabditis elegans
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DOI：
10.2144/000113365
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发表时间：
2010-03-01
期刊：
影响因子：
2.7
通讯作者：
Lee, Junho
中科院分区：
文献类型：
--
作者：
Min, Kyoengwoo;Kang, Junsu;Lee, Junho
RNA干扰（RNAi）是秀丽隐杆线虫中常用的反向遗传学方法。饲养RNAi是进行全基因组RNAi筛选的最方便和最便宜的方法。然而，据报道，通过使用各自含有一种dsRNA种类的细菌混合物饲喂RNAi来敲低两种基因（双RNAi）产生差的结果。为了克服这种低效率的问题，我们设计并测试了使用含有两个基因片段的单个RNAi构建体的双馈入RNAi方法。从三组不同基因的实验中，我们发现新的双RNAi方法始终产生显着增强的双敲低表型。本文所述的双馈RNAi方法提供了一种方法来一致地检查由C中多于一个基因的缺失引起的表型。优雅
RNA interference (RNAi) is a commonly used technique for reverse genetic approaches in Caenorhabditis elegans. Feeding RNAi is the most convenient and inexpensive method for performing genome-wide RNAi screens. However, it has been reported that knock-down of two genes (double RNAi) by feeding RNAi using a mixture of bacteria that each contained one dsRNA species produced poor results. To overcome this problem of inefficiency, we designed and tested a double feeding RNAi method using a single RNAi construct containing two gene fragments. From experiments with three different sets of genes, we found that the new double RNAi method consistently produced significantly enhanced double knock-down phenotypes. The double feeding RNAi approach described here provides a method to consistently examine phenotypes caused by depletion of more than one gene in C. elegans.