Long・term effect of systemic RNA interference on circadian clock genes in hemimetabolous insects.
Long・term effect of systemic RNA interference on circadian clock genes in hemimetabolous insects.
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系统性RNA干扰对半变态昆虫生物钟基因的长期影响。
DOI：
10.1016/j.jinsphys.2013.02.009
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发表时间：
2013
影响因子：
2.2
通讯作者：
瓜生央大
中科院分区：
文献类型：
--
作者：
Hanafusa S.;Kawaguchi T.;Umezaki Y.;Tomioka K.;Yoshii T;瓜生央大
RNA干扰(RNAi)策略能够实现基因特异性的敲除转录本，已广泛应用于昆虫研究的广泛领域，用于研究基因功能，而不涉及模式昆虫和非模式昆虫。这项技术对促进非模式昆虫的分子研究特别有益。然而，RNAi的最佳条件仍然没有被很好地理解，因为它的效率取决于物种、靶基因和实验条件。为了将RNAi技术应用于长期的实验，如时间生物学研究，RNAi的影响必须贯穿整个实验。在本研究中，我们试图确定系统RNAi的双链RNA(DsRNA)的最适浓度及其在两种不同昆虫中的有效期，这两种不同的昆虫是板栗和火虫。在这两个物种中，较高浓度的dsRNA主要产生更有效的下调测试时钟基因的mRNA水平，尽管影响取决于基因和物种。令人惊讶的是，RNAi的作用在蟋蟀和火鸡分别注射dsRNA后1-2周和1个月达到最大值，这表明RNAi的作用是缓慢但长期的。我们的研究为RNAi技术在任何长期实验中的应用提供了基础信息。
RNA interference (RNAi) strategy, which enables gene-specific knock-down of transcripts, has been spread across a wide area of insect studies for investigating gene function without regard to model and non-model insects. This technique is of particular benefit to promote molecular studies on non-model insects. However, the optimal conditions for RNAi are still not well understood because of its variable efficiency depending on the species, target genes, and experimental conditions. To apply RNAi technique to long-running experiments such as chronobiological studies, the effects of RNAi have to persist throughout the experiment. In this study, we attempted to determine the optimal concentration of double-stranded RNA (dsRNA) for systemic RNAi and its effective period in two different insect species, the cricket Gryllus bimaculatus and the firebrat Thermobia domestica. In both species, higher concentrations of dsRNA principally yielded a more efficient knock-down of mRNA levels of tested clock genes, although the effect depended on the gene and the species. Surprisingly, the effect of the RNAi reached its maximum effect 1–2weeks and 1month after the injection of dsRNA in the crickets and the firebrats, respectively, suggesting a slow but long-term effect of RNAi. Our study provides fundamental information for utilizing RNAi technique in any long-running experiment.