Materna Stefan C, Oliveri Paola
Division of Biology, California Institute of Technology, Pasadena, California 91125, USA.
Nat Protoc. 2008;3(12):1876-87. doi: 10.1038/nprot.2008.187.
Regulatory genes form large networks that are fundamental to the developmental program. The protocol presented here describes a general approach to assemble maps of gene regulatory networks (GRNs). It combines high-resolution spatio-temporal profiling of regulatory genes, strategies to perturb gene expression and quantification of perturbation effects on other genes of the network. The map of the GRN emerges by integration of these data sources and explains developmental events in terms of functional linkages between regulatory genes. This protocol has been successfully applied to regulatory processes in the sea urchin embryo, but it is generally applicable to any developmental process that relies primarily on transcriptional regulation. Unraveling the GRN for a whole tissue or organ is a challenging undertaking and, depending on the complexity, may take anywhere from months to years to complete.
调控基因形成了对发育程序至关重要的大型网络。本文介绍的方案描述了一种构建基因调控网络（GRN）图谱的通用方法。它结合了调控基因的高分辨率时空分析、干扰基因表达的策略以及对网络中其他基因干扰效应的量化。通过整合这些数据源得出GRN图谱，并根据调控基因之间的功能联系来解释发育事件。该方案已成功应用于海胆胚胎的调控过程，但它一般适用于任何主要依赖转录调控进行的发育过程。解析整个组织或器官的GRN是一项具有挑战性的工作，根据其复杂程度，可能需要数月至数年才能完成。
