The development of single cell microfluidic technologies for analysis of microRNA expression and epigenetic state with application to hematopoietic stem cells

用于分析 microRNA 表达和表观遗传状态的单细胞微流体技术的发展及其在造血干细胞中的应用

• 批准号: 413633-2012
• 负责人: Hansen, Carl
• 金额: $ 4.19万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Collaborative Health Research Projects
• 财政年份: 2012
• 资助国家: 加拿大
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=503831
• 关键词: development; single; cell; microfluidic; technologies

This project will create new and robust technology to assess the transcriptional heterogeneity of single cells at a genomic level. The prototype example to be used as a test case is the stem cell of the hematopoietic system where compelling evidence now indicates that different fate decisions are determined in part by mechanisms that are self-propagating through many self-renewal divisions. Understanding the molecular basis of these mechanisms is fundamental to current intense efforts to expand hematopoietic stem cells ex vivo, to develop safer and more effective gene therapy protocols, and to elucidate mechanisms of leukemogenesis in individual patients. The technology to be developed, validated and tested in proof-of-principle experiments on highly purified populations of normal mouse and human hematopoietic stem cells will enable microRNA (miRNA) expression and epigenomic marks to be examined on hundreds of individual cells from small samples with high fidelity and at low cost. MiRNA analyses will utilize on-chip extraction, barcoding and PCR amplification

该项目将创建新的和强大的技术，以评估单细胞在基因组水平上的转录异质性。用作测试案例的原型例子是造血系统的干细胞，现在有令人信服的证据表明，不同的命运决定部分是由通过许多自我更新分裂自我繁殖的机制决定的。了解这些机制的分子基础是目前的努力，以扩大造血干细胞体外，开发更安全，更有效的基因治疗方案，并阐明白血病发生在个别患者的机制的基础。该技术将在高度纯化的正常小鼠和人类造血干细胞群体的原理验证实验中开发、验证和测试，将使微小RNA（miRNA）的表达和表观基因组标记能够在来自小样本的数百个单个细胞上以高保真度进行检查。并且成本低。miRNA分析将利用芯片上提取、条形码和PCR扩增