Gouda Kenji, Matsunaga Yohei, Iwasaki Takashi, Kawano Tsuyoshi
Department of Bioresource Sciences, Faculty of Agriculture, Tottori University, Koyama, Japan.
Biosci Biotechnol Biochem. 2010;74(11):2361-5. doi: 10.1271/bbb.100579. Epub 2010 Nov 7.
In reverse genetics, RNA interference (RNAi) which is substitutable for gene-disruption, is an outstanding method for knockdown of a gene's function. In Caenorhabditis elegans, feeding RNAi is most convenient, but this RNAi is not suitable for knockdown of multiple genes. Hence, we attempted to establish an efficient method of feeding RNAi for multiple knockdown. We produced bacteria yielding three distinct double-stranded RNAs bound to one another, and fed those bacteria to C. elegans. Quantitative RT-PCR and observation of phenotypes indicated that our method is much more efficient than the traditional one. Our method is useful for investigating genes' functions in C. elegans.
在反向遗传学中，可替代基因破坏的RNA干扰（RNAi）是一种用于敲低基因功能的出色方法。在秀丽隐杆线虫中，喂食RNAi最为便捷，但这种RNAi不适用于多个基因的敲低。因此，我们尝试建立一种高效的用于多个基因敲低的喂食RNAi方法。我们制备了能产生三种彼此相连的不同双链RNA的细菌，并将这些细菌喂食给秀丽隐杆线虫。定量逆转录PCR和表型观察表明，我们的方法比传统方法效率更高。我们的方法对于研究秀丽隐杆线虫中基因的功能很有用。
