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中文摘要
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项目摘要 在单个细胞中分析激酶及其活性将使探索特定的细胞类型成为可能 功能和生化途径。虽然我们已经学到了很多关于转录的知识 在过去的十年里,通过单细胞基因组学,我们仍然了解到不同细胞类型的状态 对蛋白质,特别是不同细胞群体中的蛋白激酶网络的研究甚少。这在很大程度上 因为单细胞基因组分析面临着重大的技术挑战。与核酸不同, 蛋白质不能被放大,因此很难从单个细胞中检测到微量的蛋白质。我们 最近演示了一项原理验证实验,以准确检测蛋白质PTM亚型 在单细胞中。在这里,我们建议将此方法扩展到全球级别的配置文件 细胞培养和组织中的单个细胞。我们将整合单细胞基因组分析 用RNA测量来识别细胞类型特定的激酶图谱。最终,我们将绘制 在小鼠和人脑中检测激酶通路的具有空间背景的动态组 参与衰老和神经退化,这可能识别可用药的靶点。这个项目是一个 与我们目前的研究大相径庭。它的目标雄心勃勃,但是可以实现的。该项目 将利用我们在单电池技术开发和新技术应用方面的专业知识 技术应用于不同的生物系统。
英文摘要
Project Summary Profiling kinases and their activities in single cells will enable exploration of cell type specific functional and biochemical pathways. While we have learned a great deal about transcriptional states of different cell types in the past decade through single cell genomics, we still understand little about proteins and in particular kinases networks in diverse cell populations. This is largely because single-cell kinome analysis faces major technical challenges. Unlike nucleic acids, proteins cannot be amplified, making detection of minute quantities from single cells difficult. We recently demonstrated a proof-of-principle experiment to accurately detect protein PTM isoforms in single cells. Here we propose to scale this method up to profile kinases at the global level in single cells in both cell culture and in tissues. We will integrate the single cell kinome analysis with RNA measurements to identify cell type specific kinase profiles. Ultimately, we will map the kinome with spatial context in the mouse and human brain to examine the kinase pathways involved in aging and neurodegeneration which could identify druggable targets. This project is a major departure from our current research. Its goals are ambitious but achievable. The project will leverage our expertise in single cell technology development and in application of new technology to diverse biological systems.
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Single cell analysis of the kinome
A regulome and transcriptome atlas of fetal and adult human neurogenesis
Dynamics of chromosome organization and chromatin states in single cells
Dynamics of chromosome organization and chromatin states in single cells