Determination of absolute expression profiles using multiplexed miRNA analysis.
Determination of absolute expression profiles using multiplexed miRNA analysis.
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DOI：
10.1371/journal.pone.0180988
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Wang TH
中科院分区：
文献类型：
--
作者：
Song Y;Kilburn D;Song JH;Cheng Y;Saeui CT;Cheung DG;Croce CM;Yarema KJ;Meltzer SJ;Liu KJ;Wang TH
准确测量miRNA的表达对于了解它们在基因表达中的作用以及作为疾病生物标记物的应用至关重要。正确识别miRNA表达的变化依赖于可靠的正常化，以解释样本之间的生物和技术差异。LIGO-miR是一种多重分析方法，旨在快速测量miRNA的绝对拷贝数，从而减少对生物控制的依赖。它使用简单的两步连接过程来产生长度编码的产物，这些编码产物可以使用各种DNA大小方法进行量化。我们展示了Ligo-miR能够将miRNA表达量化到每个细胞20个拷贝的敏感度，准确区分密切相关的miRNA，并可靠地测量小至1.2倍的差异变化。然后，进行标杆研究，以显示Ligo-miR、微阵列和TaqMan qRT-PCR之间的高度相关性。最后，Ligo-miR被用来确定一些乳腺、食道和胰腺细胞系中的拷贝数图谱，并展示了拷贝数分析在提供对表达谱变化的分层洞察方面的实用性。
Accurate measurement of miRNA expression is critical to understanding their role in gene expression as well as their application as disease biomarkers. Correct identification of changes in miRNA expression rests on reliable normalization to account for biological and technological variance between samples. Ligo-miR is a multiplex assay designed to rapidly measure absolute miRNA copy numbers, thus reducing dependence on biological controls. It uses a simple 2-step ligation process to generate length coded products that can be quantified using a variety of DNA sizing methods. We demonstrate Ligo-miR’s ability to quantify miRNA expression down to 20 copies per cell sensitivity, accurately discriminate between closely related miRNA, and reliably measure differential changes as small as 1.2-fold. Then, benchmarking studies were performed to show the high correlation between Ligo-miR, microarray, and TaqMan qRT-PCR. Finally, Ligo-miR was used to determine copy number profiles in a number of breast, esophageal, and pancreatic cell lines and to demonstrate the utility of copy number analysis for providing layered insight into expression profile changes.
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影响因子：
15
作者：
Liu, Kelvin J.;Rane, Tushar D.;Zhang, Yi;Wang, Tza-Huei
通讯作者：
Wang, Tza-Huei
影响因子：
14.9
作者：
Chen C;Ridzon DA;Broomer AJ;Zhou Z;Lee DH;Nguyen JT;Barbisin M;Xu NL;Mahuvakar VR;Andersen MR;Lao KQ;Livak KJ;Guegler KJ
通讯作者：
Guegler KJ
DOI：
10.1073/pnas.0800121105
发表时间：
2008-04-01
影响因子：
11.1
作者：
Calin, George A.;Cimmino, Amelia;Croce, Carlo M.
通讯作者：
Croce, Carlo M.
影响因子：
4.5
作者：
Burgos, Kasandra Lovette;Javaherian, Ashkan;Van Keuren-Jensen, Kendall
通讯作者：
Van Keuren-Jensen, Kendall
影响因子：
82.9
作者：
Bonci, Desiree;Coppola, Valeria;De Maria, Ruggero
通讯作者：
De Maria, Ruggero