Precise mapping of the transcription start sites of human microRNAs using DROSHA knockout cells.
Precise mapping of the transcription start sites of human microRNAs using DROSHA knockout cells.
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DOI：
10.1186/s12864-016-3252-7
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发表时间：
2016-11-11
期刊：
影响因子：
4.4
通讯作者：
Kim YK
中科院分区：
文献类型：
--
作者：
Jeong G;Lim YH;Kim YK
MicroRNAs(MiRNAs)的表达主要在其转录过程中受到调控。然而，由于对大多数miRNAs的启动子和转录起始点缺乏足够的信息，miRNA基因的转录调控还没有得到广泛的研究。在本研究中，我们使用DROSHA基因敲除细胞鉴定了人类初级miRNAs(pri-miRNAs)的转录起始位置。DROSHA基因敲除导致pri-miRNAs的积累增加，并有助于利用快速扩增cDNA末端(RACE)技术精确定位其5‘端核苷酸。通过分析miRNAs转录起始点周围的启动子区域，我们发现它们序列中不相关的miRNAs在其启动子中有许多共同的元件来结合相同的转录因子。此外，通过分析内含子miRNAs，我们还获得了全面的证据，表明含有miRNA的内含子拼接速度比其他内含子慢。精确定位的pri-miRNAs转录起始点，以及pri-miRNAs调控的转录因子列表，将为未来研究了解miRNAs的调控网络提供宝贵的资源。本文的在线版本(doi：10.1186/s12864-0163252-7)包含补充材料，授权用户可以使用。
The expression of microRNAs (miRNAs) is primarily regulated during their transcription. However, the transcriptional regulation of miRNA genes has not been studied extensively owing to the lack of sufficient information about the promoters and transcription start sites of most miRNAs. In this study, we identified the transcription start sites of human primary miRNAs (pri-miRNAs) using DROSHA knockout cells. DROSHA knockout resulted in increased accumulation of pri-miRNAs and facilitated the precise mapping of their 5′ end nucleotides using the rapid amplification of cDNA ends (RACE) technique. By analyzing the promoter region encompassing the transcription start sites of miRNAs, we found that the unrelated miRNAs in their sequences have many common elements in their promoters for binding the same transcription factors. Moreover, by analyzing intronic miRNAs, we also obtained comprehensive evidence that miRNA-harboring introns are spliced more slowly than other introns. The precisely mapped transcription start sites of pri-miRNAs, and the list of transcription factors for pri-miRNAs regulation, will be valuable resources for future studies to understand the regulatory network of miRNAs. The online version of this article (doi:10.1186/s12864-016-3252-7) contains supplementary material, which is available to authorized users.
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