Small RNA Sequencing across Diverse Biofluids Identifies Optimal Methods for exRNA Isolation
Small RNA Sequencing across Diverse Biofluids Identifies Optimal Methods for exRNA Isolation
复制标题
小RNA测序跨越不同的生物流体确定最佳方法的exRNA分离
DOI：
10.1016/j.cell.2019.03.024
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发表时间：
2019-04-04
期刊：
影响因子：
64.5
通讯作者：
Laurent, Louise C.
中科院分区：
文献类型：
--
作者：
Srinivasan, Srimeenakshi;Yeri, Ashish;Laurent, Louise C.
由于缺乏对细胞外RNA(ExRNA)分离方法性能的了解，研究内部和研究之间的重复性较差，阻碍了exRNA领域的进展。对5种生物体液中10种外源RNA分离方法的系统比较表明，所得到的小RNA-SEQ图谱在复杂性和重复性方面存在显著差异。通过估计每个图谱中细胞外小泡(EV)、核糖核蛋白(RNP)和高密度脂蛋白(HDL)特异的miRNA签名的比例，确定每种方法访问不同exRNA载体亚类的相对效率。开发了一个基于网络的交互式应用程序(MiRDaR)，以帮助研究人员为他们的研究选择最佳的exRNA提取方法。MiRDar为每种exRNA分离方法提供所需生物液中所有表达的miRNAs或选定的miRNAs子集的比较统计数据，并返回按复杂性、表达水平和重复性优先排序的exRNA分离方法的排序列表。这些结果将改善重复性，并刺激外源RNA生物标记物的进一步发展。R为每种exRNA分离方法提供所需生物液中所有表达的miRNAs或选定的miRNAs子集的比较统计数据，并返回按复杂性、表达水平和重复性优先排序的exRNA分离方法的排序列表。这些结果将改善重复性，并刺激外源RNA生物标记物开发的进一步进展。
Poor reproducibility within and across studies arising from lack of knowledge regarding the performance of extracellular RNA (exRNA) isolation methods has hindered progress in the exRNA field. A systematic comparison of 10 exRNA isolation methods across 5 biofluids revealed marked differences in the complexity and reproducibility of the resulting small RNA-seq pro- files. The relative efficiency with which each method accessed different exRNA carrier subclasses was determined by estimating the proportions of extracellular vesicle (EV)-, ribonucleoprotein (RNP)-, and highdensity lipoprotein (HDL)-specific miRNA signatures in each profile. An interactive web-based application (miRDaR) was developed to help investigators select the optimal exRNA isolation method for their studies. miRDar provides comparative statistics for all expressed miRNAs or a selected subset of miRNAs in the desired biofluid for each exRNA isolation method and returns a ranked list of exRNA isolation methods prioritized by complexity, expression level, and repro ducibility. These results will improve reproducibility and stimulate further progret in exRNA biomarker development. r provides comparative statistics for all expressed miRNAs or a selected subset of miRNAs in the desired biofluid for each exRNA isolation method and returns a ranked list of exRNA isolation methods prioritized by complexity, expression level, and repro ducibility. These results will improve reproducibility and stimulate further progress in exRNA biomarker development.