Journal
BIOTECHNIQUES
Volume 48, Issue 3, Pages 229-232Publisher
FUTURE SCI LTD
DOI: 10.2144/000113365
Keywords
C. elegans; feeding RNA interference; double knock-down
Funding
- Research Center for Functional Cellulomics at Seoul National University
- Korea Science and Engineering Foundation (KOSEF)
- Ministry of Science and Technology [20090083336]
- Ministry for Health, Welfare and Family Affairs [0920230]
- Korean Research Foundation [2005-037-C00027]
- Brain Korea 21 (BK21) program [2007]
- Seoul RBD Program [2007]
- Korea Health Promotion Institute [0920230] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- National Research Foundation of Korea [2008-0057809, 2005-037-C00027, 2007-03651] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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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.
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