利用自动化单细胞分析技术解析线虫进化保守基因的发育功能与调控网络

基因组如何调控复杂发育是遗传学与发育生物学研究的重大课题。高通量遗传干扰、高分辨率活体成像、丰富的荧光标记资源和基于图像处理的单细胞分析手段的发展为大规模、高通量地解析基因组的发育功能和调控网络提供契机。本课题拟以线虫胚胎发育为模型，应用活体成像和自动单细胞表型分析等最新方法解析进化保守的基因及其网络如何调控胚胎发育。本课题拟（1）对1,000个进化保守的调控基因进行单基因失活干扰，利用活体显微成像和自动化单细胞表型分析方法，在细胞水平测定细胞增殖、分化和形态建成等发育表型；（2）通过对表型的生物信息分析鉴定基因在特定细胞和发育阶段的调控功能；（3）构建多层次发育调控网络（iDevNet）探索网络的基本特征与规律及保守基因的功能模式。（4）构建发育数据库（iDevBase）分享表型与功能数据，为数量发育生物学研究提供重要资源与推动。本课题对理解基因组功能、发育过程和调控网络有重要意义。

How genome regulates complex in vivo developmental processes is a central question in genetics and developmental biology. Recent progresses in high-throughput genetic interference, high-resolution live imaging, large-scale fluorescent reagents and high-accuracy image processing-based single-cell analyses enable systems-level analysis of genomic regulation of in vivo development. We propose to use C. elegans embryogenesis as a model to investigate how the evolutionarily conserved genes and their networks regulate complex in vivo development using live imaging and automated single-cell analyses. We plan to (1) perturb 1,000 highly conserved genes individually and quantify cellular developmental phenotypes in terms of proliferation, differentiation and morphogenesis using a combination of live imaging and automated single-cell phenotyping; (2) infer each gene’s function in specific cells and developmental stages through systematic phenotypic analysis; (3) construct an integrated developmental network (iDevNet) to explore general properties and principles of developmental regulatory network; and (4) construct a comprehensive open-access developmental biology database (iDevBase) for quantitative developmental biology, including images, cellular phenotypic measurements, inferred gene function and integrated network. The significance of this proposal is twofold: first, it will reveal new gene function, novel regulatory processes and integrated networks of in vivo development. In addition, it will provide a valuable resource in terms of the richness of information, resolution and scale for follow-up mechanistic analyses, network simulation and systems biology of development.