Ingestion of bacterially expressed dsRNAs can produce specific and potent genetic interference in Caenorhabditis elegans
Ingestion of bacterially expressed dsRNAs can produce specific and potent genetic interference in Caenorhabditis elegans
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DOI：
10.1016/s0378-1119(00)00579-5
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发表时间：
2001-01-24
期刊：
影响因子：
3.5
通讯作者：
Fire, A
中科院分区：
文献类型：
--
作者：
Timmons, L;Court, DL;Fire, A
双链RNA(RNAi)介导的遗传干扰已成为分析秀丽线虫基因功能的重要工具。在这里，我们报告了一种使用细菌传递双链RNA的有效诱导RNAi的方法。这种方法利用了缺乏RNaseIII的细菌，RNaseIII是一种通常能降解细菌细胞中大多数dsRNA的酶。缺乏RNaseIII的细菌被改造成产生大量特定的dsRNA片段。当喂给线虫时，这些工程菌被发现产生了受RNAi影响的动物群体，其表型在表达能力上与相应的功能丧失突变体相当。我们发现这种方法在诱导晚期幼虫和成虫两性非神经元组织的RNAi方面最有效，但在神经系统、早期幼虫阶段和雄性中的有效性降低。细菌诱导的RNAi表型可以在连续饲养的几代过程中保持不变，从而能够方便地评估特定遗传干扰和持续暴露于dsRNAs的生物学后果。(C)2001 Elsevier Science B.V.保留所有权利。
Genetic interference mediated by double-stranded RNA (RNAi) has been a valuable tool in the analysis of gene function in Caenorhabditis elegans. Here we report an efficient induction of RNAi using bacteria to deliver double-stranded RNA. This method makes use of bacteria that are deficient in RNaseIII, an enzyme that normally degrades a majority of dsRNAs in the bacterial cell. Bacteria deficient for RNaseIII were engineered to produce high quantities of specific dsRNA segments. When fed to C. elegans, such engineered bacteria were found to produce populations of RNAi-affected animals with phenotypes that were comparable in expressivity to the corresponding loss-of-function mutants. We found the method to be most effective in inducing RNAi for non-neuronal tissue of late larval and adult hermaphrodites, with decreased effectiveness in the nervous system, in early larval stages, and in males. Bacteria-induced RNAi phenotypes could be maintained over the course of several generations with continuous feeding, allowing for convenient assessments of the biological consequences of specific genetic interference and of continuous exposure to dsRNAs. (C) 2001 Elsevier Science B.V. All rights reserved.