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在高时间分辨率下测量绝对RNA拷贝数揭示发育过程中的转录组动力学。

Measuring Absolute RNA Copy Numbers at High Temporal Resolution Reveals Transcriptome Kinetics in Development.

作者信息

Owens Nick D L, Blitz Ira L, Lane Maura A, Patrushev Ilya, Overton John D, Gilchrist Michael J, Cho Ken W Y, Khokha Mustafa K

机构信息

The Francis Crick Institute, Mill Hill Laboratory, The Ridgeway Mill Hill, London NW7 1AA, UK.

Department of Developmental and Cell Biology, University of California, Irvine, CA 92697 USA.

出版信息

Cell Rep. 2016 Jan 26;14(3):632-647. doi: 10.1016/j.celrep.2015.12.050. Epub 2016 Jan 7.

Abstract

Transcript regulation is essential for cell function, and misregulation can lead to disease. Despite technologies to survey the transcriptome, we lack a comprehensive understanding of transcript kinetics, which limits quantitative biology. This is an acute challenge in embryonic development, where rapid changes in gene expression dictate cell fate decisions. By ultra-high-frequency sampling of Xenopus embryos and absolute normalization of sequence reads, we present smooth gene expression trajectories in absolute transcript numbers. During a developmental period approximating the first 8 weeks of human gestation, transcript kinetics vary by eight orders of magnitude. Ordering genes by expression dynamics, we find that "temporal synexpression" predicts common gene function. Remarkably, a single parameter, the characteristic timescale, can classify transcript kinetics globally and distinguish genes regulating development from those involved in cellular metabolism. Overall, our analysis provides unprecedented insight into the reorganization of maternal and embryonic transcripts and redefines our ability to perform quantitative biology.

摘要

转录调控对细胞功能至关重要,调控异常会导致疾病。尽管有技术可用于检测转录组,但我们对转录动力学缺乏全面的了解,这限制了定量生物学的发展。这在胚胎发育中是一个严峻的挑战,因为基因表达的快速变化决定了细胞命运的抉择。通过对非洲爪蟾胚胎进行超高频采样以及对序列读数进行绝对标准化,我们展示了以绝对转录本数量表示的平滑基因表达轨迹。在近似人类妊娠前8周的发育阶段,转录动力学变化幅度达八个数量级。根据表达动态对基因进行排序,我们发现“时间共表达”可预测常见的基因功能。值得注意的是,一个单一参数,即特征时间尺度,能够全局分类转录动力学,并区分调控发育的基因与参与细胞代谢的基因。总体而言,我们的分析为母体和胚胎转录本的重组提供了前所未有的见解,并重新定义了我们进行定量生物学研究的能力。

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