一种改良的喂食 RNAi 方法，可用于同时敲低秀丽隐杆线虫中多个基因。

A modified feeding RNAi method for simultaneous knock-down of more than one gene in Caenorhabditis elegans.

机构信息

Research Center for Functional Cellulomics, Institute of Molecular Biology and Genetics, School of Biological Sciences, Seoul National University, Seoul, Korea.

出版信息

Biotechniques. 2010 Mar;48(3):229-32. doi: 10.2144/000113365.

DOI:10.2144/000113365

PMID:20359305

Abstract

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.

摘要

RNA 干扰（RNAi）是一种在秀丽隐杆线虫中常用的反向遗传学方法。喂食 RNAi 是进行全基因组 RNAi 筛选最方便和最便宜的方法。然而，据报道，使用含有两种 dsRNA 物种的细菌混合物进行喂食 RNAi 会导致两个基因的敲低（双 RNAi）效果不佳。为了克服这种效率低下的问题，我们设计并测试了一种使用含有两个基因片段的单个 RNAi 构建体的双喂食 RNAi 方法。通过对三组不同基因的实验，我们发现新的双 RNAi 方法始终产生显著增强的双敲低表型。这里描述的双喂食 RNAi 方法为在秀丽隐杆线虫中研究多个基因缺失引起的表型提供了一种方法。