C. elegans rrf-1 mutations maintain RNAi efficiency in the soma in addition to the germline.
C. elegans rrf-1 mutations maintain RNAi efficiency in the soma in addition to the germline.
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DOI：
10.1371/journal.pone.0035428
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Hansen M
中科院分区：
文献类型：
--
作者：
Kumsta C;Hansen M
通过RNA干扰（RNAi）使基因失活已被证明是研究C.优美的当与组织特异性基因失活方法相结合时，RNAi有可能揭示基因在不同组织中的功能。在这项研究中，我们确定了C。线虫rrf-1突变体，以确定它们在各种组织中处理RNAi的能力。这些突变体已被广泛用于RNAi研究中，以评估C.秀丽隐杆线虫生殖系经过进一步分析，我们发现两个携带不同功能丧失等位基因的rrf-1突变体能够处理针对几个体细胞表达基因的RNAi。具体来说，我们观察到肠道能够有效地处理RNAi触发物，而皮下组织中的细胞对rrf-1突变体中的RNAi表现出部分易感性。rrf-1突变体中的其他体细胞组织，如肌肉和体细胞性腺，似乎对RNAi具有抗性。除了这些观察结果，我们发现rrf-1（pk 1417）突变诱导了几个转基因阵列的表达，包括FOXO转录因子β 16。出乎意料的是，rrf-1（pk 1417）突变体表现出增加的内源性表达的pk 16靶基因sod-3;然而，rrf-1（pk 1417）突变体的寿命和耐热性与野生型动物相似。总之，这些数据表明，rrf-1突变体显示了几种以前没有认识到的表型，包括更广泛的组织特异性RNAi加工能力，我们的研究结果强调了需要仔细表征组织特异性RNAi工具。
Gene inactivation through RNA interference (RNAi) has proven to be a valuable tool for studying gene function in C. elegans. When combined with tissue-specific gene inactivation methods, RNAi has the potential to shed light on the function of a gene in distinct tissues. In this study we characterized C. elegans rrf-1 mutants to determine their ability to process RNAi in various tissues. These mutants have been widely used in RNAi studies to assess the function of genes specifically in the C. elegans germline. Upon closer analysis, we found that two rrf-1 mutants carrying different loss-of-function alleles were capable of processing RNAi targeting several somatically expressed genes. Specifically, we observed that the intestine was able to process RNAi triggers efficiently, whereas cells in the hypodermis showed partial susceptibility to RNAi in rrf-1 mutants. Other somatic tissues in rrf-1 mutants, such as the muscles and the somatic gonad, appeared resistant to RNAi. In addition to these observations, we found that the rrf-1(pk1417) mutation induced the expression of several transgenic arrays, including the FOXO transcription factor DAF-16. Unexpectedly, rrf-1(pk1417) mutants showed increased endogenous expression of the DAF-16 target gene sod-3; however, the lifespan and thermo-tolerance of rrf-1(pk1417) mutants were similar to those of wild-type animals. In sum, these data show that rrf-1 mutants display several phenotypes not previously appreciated, including broader tissue-specific RNAi-processing capabilities, and our results underscore the need for careful characterization of tissue-specific RNAi tools.
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