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RNA干扰:哺乳动物卵母细胞无需依赖RNA的RNA聚合酶即可实现RNA干扰。

RNAi: mammalian oocytes do it without RNA-dependent RNA polymerase.

作者信息

Stein Paula, Svoboda Petr, Anger Martin, Schultz Richard M

机构信息

Department of Biology, University of Pennsylvania, 415 South University Avenue, Philadelphia, PA 19104-6018, USA.

出版信息

RNA. 2003 Feb;9(2):187-92. doi: 10.1261/rna.2860603.

Abstract

Studies in mutant organisms deficient in RNA interference (RNAi) and related post-transcriptional gene silencing implicated a role for a single class of RNA-dependent RNA polymerases (RdRp). Nevertheless, sequence homologs to these RdRps have not been found in coelomate organisms such as Drosophila or mammals. This lack of homologous sequences does not exclude that an RdRp functions in RNAi in these organisms because an RdRp could be acquired by horizontal transfer from an RNA virus. In fact, such a sequence is found in mice (Aquarius) and we observe that it is expressed in mouse oocytes and early embryos, which exhibit RNAi. We report here that cordycepin, an inhibitor of RNA synthesis, does not prevent Mos double-strand RNA (dsRNA) to target endogenous Mos mRNA in mouse oocytes and that targeting a chimeric Mos-EGFP mRNA with dsRNA to EGFP does not reduce the endogenous Mos mRNA, but does target the chimeric mRNA. These results indicate that an RdRp is not involved in dsRNA-mediated mRNA degradation in mammalian oocytes, and possibly in mammals in general, and therefore that only homologous sequences to the dsRNA are targeted for degradation.

摘要

对缺乏RNA干扰(RNAi)及相关转录后基因沉默的突变生物体的研究表明,一类RNA依赖性RNA聚合酶(RdRp)发挥了作用。然而,在果蝇或哺乳动物等真体腔生物中尚未发现与这些RdRp的序列同源物。缺乏同源序列并不排除RdRp在这些生物体的RNAi中发挥作用,因为RdRp可能通过水平转移从RNA病毒获得。事实上,在小鼠中发现了这样一个序列(水瓶座蛋白),并且我们观察到它在表现出RNAi的小鼠卵母细胞和早期胚胎中表达。我们在此报告,RNA合成抑制剂虫草素并不能阻止双链RNA(dsRNA)靶向小鼠卵母细胞中的内源性Mos mRNA,并且用针对EGFP的dsRNA靶向嵌合的Mos-EGFP mRNA并不会降低内源性Mos mRNA,但确实会靶向嵌合mRNA。这些结果表明,RdRp不参与哺乳动物卵母细胞中dsRNA介导的mRNA降解,可能在一般哺乳动物中也是如此,因此只有与dsRNA的同源序列被靶向降解。

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