High-throughput single-cell analysis in microfluidically-generated droplets

微流体产生的液滴的高通量单细胞分析

• 批准号: 9552549
• 负责人: Eli Boritz
• 金额: $ 56.97万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9552549
• 关键词: Address; Biological; Cells; Complex; Emerging Technologies; Emulsions; Encapsulated; Genome; Heterogeneity; Immunology; Individual; Laboratories; Libraries; Microfluidics; Nucleic Acids; Population; Preparation; Reaction; Resolution; Signal Transduction; Technology; Virus; Work; cell population study; method development; next generation sequencing; single cell analysis; transcriptome

This project was initiated at the end of the fiscal year in a newly-created section of the Immunology Laboratory, VRC to leverage work using microfluidics undertaken to answer a specific biological question in a separate project. The overarching rationale for this project is that emerging technologies for next-generation sequencing allow the comprehensive determination of genomes, transcriptomes, and other complex populations of nucleic acid, but on their own do not permit resolution of signal from individual cells within complex populations. This is a significant limitation in multiple settings where information about intercellular heterogeneity is needed. These include the analysis of rare virus-infected cells in large populations as well as the global understanding of highly diverse and complex populations. To address this need, this project seeks to employ microfluidics to generate stable, uniform emulsions in which single cells are encapsulated at limiting dilution. Thus, tens of millions of single-cell reaction chambers can be created. Various adaptations of next-generation sequencing library preparation technology are then employed to generate material for sequencing that capture information about intercellular heterogeneity within cell populations studied.

该项目于本财年末在 VRC 免疫学实验室新创建的部门启动，旨在利用微流体技术在一个单独的项目中回答特定的生物学问题。 该项目的总体原理是，新一代测序的新兴技术可以全面测定基因组、转录组和其他复杂的核酸群体，但其本身并不能解析复杂群体中单个细胞的信号。 这在需要细胞间异质性信息的多种环境中是一个显着的限制。 其中包括对大量人群中罕见病毒感染细胞的分析，以及对高度多样化和复杂人群的全球了解。 为了满足这一需求，该项目寻求采用微流体技术来生成稳定、均匀的乳液，其中单细胞在有限稀释下被封装。 因此，可以创建数千万个单细胞反应室。 然后采用下一代测序文库制备技术的各种改进来生成用于测序的材料，以捕获有关所研究的细胞群内的细胞间异质性的信息。