A modified feeding RNAi method for simultaneous knock-down of more than one gene in Caenorhabditis elegans.
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.
Min
Kyoengwoo
K
Research Center for Functional Cellulomics, Institute of Molecular Biology and Genetics, School of Biological Sciences, Seoul National University, Seoul, Korea.
Kang
Junsu
J
Lee
Junho
J
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics*; D055785:Gene Knockdown Techniques / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D010641:Phenotype; D034622:RNA Interference
D029742:Caenorhabditis elegans Proteins