dRNA-seq原理及其在原核生物转录组学研究中的应用
收稿日期: 2013-03-20
修回日期: 2013-05-16
网络出版日期: 2013-08-25
基金资助
国家自然科学基金项目(编号：31270132)资助
The principle of dRNA-seq and its applications in prokaryotic tran-scriptome analyses
Received date: 2013-03-20
Revised date: 2013-05-16
Online published: 2013-08-25
第二代测序技术不仅推进了基因组学的研究, 而且也广泛应用到转录组学的研究中。原核生物转录组学的研究方法主要有Tiling芯片和RNA-seq, 后者因其较高的覆盖率和分辨率以及越来越低廉的成本而逐渐被更多的研究单位采用。根据对mRNA富集方法或rRNA去除方法的不同, 目前RNA-seq方法主要有6种, 其中dRNA-seq由于采用了5′单磷酸依赖的外切酶, 可以特异性地降解加工过的转录本, 从而使初始转录本得到富集, 已被广泛应用到原核生物转录起始位点、小的调控RNA、启动子及操纵子等研究中。文章对dRNA-seq的原理、技术流程及其在原核生物转录组学研究中的应用进行了综述, 并对这一研究方法在现阶段应用的优势和局限性进行了讨论, 也进一步对该方法在未来的发展和应用进行了展望, 期望为国内相关领域研究人员提供参考。
侯志伟 王赟 高宏 侯圣伟 . dRNA-seq原理及其在原核生物转录组学研究中的应用[J]. 遗传, 2013 , 35(8) : 983 -991 . DOI: 10.3724/SP.J.1005.2013.00983
The genomic and transcriptomic studies have been greatly accelerated by the next-generation sequencing technology. Currently, there are two main methods in transcriptomic studies, namely, Tiling microarray and RNA-seq. Comparatively, because of its overwhelming superiority in transcript coverage and resolution, as well as decrease of costs, RNA-seq has been employed in transcriptome analyses by an increasing number of research institutes. According to the mRNA enrichment or rRNA depletion methods, RNA-seq can be performed in six ways. Among them, dRNA-seq can descriminate the original transcripts from the processed RNAs based on a differential exonuclease treatment. This kind of method has been broadly applied in studies of prokaryotic transcriptional start sites, small regulatory RNAs, promoters and operons, etc. Here, we reviewed the detailed principle and technical process of dRNA-seq and its applications in prokaryotic transcriptome analyses. Besides, we also summarized the advantages and limitations of dRNA-seq at present stage, and then gave our perspective of its future development and potential applications, expecting to present useful references for civil researchers in related fields.
Key words： dRNA-seq; sRNA; transcriptome sequencing; TSS; ncRNA
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