Automated single-cell gene expression and phenotype analysis in C. elegans

总结： 生物体的基因组如何指导其发育和行为才刚刚开始被理解。 了解每个细胞中哪些基因起作用将是迈向全面研究的重要一步。 认识我们已经开发了一个系统，利用不变的血统的线虫C。elegans 在单细胞水平上自动定义整个发育过程中的基因表达和表型， 高时间分辨率。该系统使用2色，3D，延时共聚焦显微镜记录 胚胎发生，然后进行自动图像分析，通过 运动、分裂和死亡。到目前为止，它可以通过十轮中的前九轮来追踪血统。 胚胎细胞分裂的高精度和分配表达的个别细胞。在这里，我们建议 系统的进一步发展。我们将通过最后一轮细胞分裂来扩展系统。我们将 还通过测量各种时间和空间特征使系统适于自动化表型分析 以及基因表达的变化。此外，我们将使用这些方法来 系统地解码已知RNAi表型的转录因子和其他基因的功能 胚胎致死 相关性： 完成的系统将是有用的各种功能基因组学的应用， 为全面阐明C.优雅等 知识在C。秀丽线虫不仅可以提供对动物发育中基因和网络的深入了解， 这表明同源基因在人类健康和疾病中的作用。

Summary: How an organism's genome directs its development and behavior is only beginning to be understood. Knowledge of what genes function in each cell would be a significant step toward a comprehensive understanding. We have developed a system that exploits the invariant lineage of the nematode C. elegans to automatically define gene expression and phenotype throughout development at the single cell level with high temporal resolution. The system uses 2-color, 3D, time-lapse confocal microscopy to record embryogenesis, followed by automated image analysis that identifies and tracks each nucleus through movement, division and death. To date, it can trace the lineage through the first nine out of ten rounds of embryonic cell divisions with high accuracy and assign expression to individual cells. Here, we propose further development of the system. We will extend the system through the last round of cell division. We will also adapt the system for automated phenotypic analysis by measuring various temporal and spatial features of cells and cell groups, as well as changes of gene expression. Furthermore, we will use the methods to systematically decode the function of transcription factors and other genes whose RNAi phenotype is known to be embryonic lethal. Relevance: The completed system will be useful for a wide variety of functional genomics applications and would provide an avenue to comprehensive elucidation of gene function and regulation in C. elegans. Such knowledge in C. elegans would not only provide insights into genes and networks in animal development, but would suggest roles of homologous genes in human health and disease.