Materna Stefan C, Nam Jongmin, Davidson Eric H
Division of Biology 156-29, California Institute of Technology, Pasadena, CA 91125, USA.
Gene Expr Patterns. 2010 Jun;10(4-5):177-84. doi: 10.1016/j.gep.2010.04.002. Epub 2010 Apr 14.
Accurate measurements of transcript abundance are a prerequisite to understand gene activity in development. Using the NanoString nCounter, an RNA counting device, we measured the prevalence of 172 transcription factors and signaling molecules in early sea urchin development. These measurements show high fidelity over more than five orders of magnitude down to a few transcripts per embryo. Most of the genes included are locally restricted in their spatial expression, and contribute to the divergent regulatory states of cells in the developing embryo. In order to obtain high-resolution expression profiles from fertilization to late gastrulation samples were collected at hourly intervals. The measured time courses agree well with, and substantially extend, prior relative abundance measurements obtained by quantitative PCR. High temporal resolution permits sequences of successively activated genes to be precisely delineated providing an ancillary tool for assembling maps of gene regulatory networks. The data are available via an interactive website for quick plotting of selected time courses.
准确测量转录本丰度是理解发育过程中基因活性的先决条件。我们使用RNA计数设备NanoString nCounter,测量了172种转录因子和信号分子在海胆早期发育中的丰度。这些测量在超过五个数量级的范围内具有高保真度,低至每个胚胎仅有少数转录本。所包含的大多数基因在空间表达上受到局部限制,并促成发育中胚胎细胞不同的调控状态。为了获得从受精到原肠胚晚期的高分辨率表达谱,每隔一小时收集一次样本。所测量的时间进程与通过定量PCR获得的先前相对丰度测量结果吻合良好,并大大扩展了这些结果。高时间分辨率使得能够精确描绘相继激活的基因序列,为构建基因调控网络图谱提供了辅助工具。这些数据可通过一个交互式网站获取,以便快速绘制选定的时间进程。