Genetic analysis of single cells

单细胞遗传分析

• 批准号: 386152-2010
• 负责人: Hansen, Carl
• 金额: $ 4.44万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2011
• 资助国家: 加拿大
• 起止时间: 2011-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=485199
• 关键词: Genetic; analysis; single; cells

A fundamental and unmet challenge in biological research is the need to understand and measure the genome and transcriptional state of individual cells. For instance, nearly all microorganisms on the planet have not been cultivated and may very well be refractory to culture in laboratory settings due to requisite symbiotic relationships with other organisms. Thus, these life forms are not amenable to current technologies for genomic study, which require large numbers and pure populations of cells. On the other hand, even genetically identical cells, such as those in the human body, exhibit widespread heterogeneity owing to differentiation state, stochastic variations, and disease. Scalable single cell analysis technologies are urgently needed and will have far reaching impact touching diverse fields including microbial diversity, cancer biology, and human development. Although advances in instrumentation and chemistries for global sequencing and sensitive targeted measurements of transcription are now available, these are limited by difficulties in manipulating single cells and inefficiencies arising from conventional large-volume (µL) sample processing. Here we will couple the power of these established technologies with new engineering methods for the fabrication of highly integrated "lab-on-a-chip" devices to develop microsystems capable of processing hundreds to thousands of single cells with improved sensitivity and speed. Specifically, we will develop two technologies for i) the high-throughput isolation and genome amplification of uncultivated microbes derived from environmental samples, and ii) the scalable analysis of miRNA expression in mammalian cells.

生物学研究中一个基本且尚未解决的挑战是需要了解和测量单个细胞的基因组和转录状态。 例如，地球上几乎所有微生物都没有经过培养，并且由于与其他生物体存在必要的共生关系，很可能难以在实验室环境中培养。 因此，这些生命形式不适合当前的基因组研究技术，因为这需要大量和纯的细胞群。 另一方面，即使是基因相同的细胞，例如人体中的细胞，由于分化状态、随机变异和疾病，也表现出广泛的异质性。迫切需要可扩展的单细胞分析技术，并将对微生物多样性、癌症生物学和人类发展等不同领域产生深远影响。尽管目前用于全局测序和灵敏的转录目标测量的仪器和化学技术已经取得了进步，但这些技术仍受到单细胞操作困难以及传统大体积 (μL) 样品处理效率低下的限制。在这里，我们将把这些现有技术的力量与新的工程方法结合起来，用于制造高度集成的“芯片实验室”设备，以开发能够处理数百至数千个单细胞的微系统，并提高灵敏度和速度。具体来说，我们将开发两项技术：i）来自环境样本的未培养微生物的高通量分离和基因组扩增，以及 ii）哺乳动物细胞中 miRNA 表达的可扩展分析。