Single Cell Core

单细胞核心

基本信息

批准号：
10594524
负责人：
Peter Alan Sims
金额：
$ 37.45万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-01-01 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10594524
关键词：
2019-nCoV Activities of Daily Living Address Adult Age Antibodies Antigens B-Cell Antigen Receptor Bar Codes Cell Count Cell Separation Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing Circulation Cytomegalovirus DNA Epitopes Flow Cytometry Funding Genomic Library Genomics Human Immune Immune response Immunologic Receptors Immunology Immunophenotyping Individual Infant Influenza Joints Libraries Link Maps Membrane Proteins Messenger RNA Microfluidics Organ Donor Phenotype Population Preparation Proteome Protocols documentation Quality Control Reporting Research Personnel Resolution Resources Rest SARS-CoV-2 antigen Sampling Site Sorting Specificity Stains Surface T cell receptor repertoire sequencing T-Cell Activation T-Cell Antigen Receptor Specificity T-Cell Receptor T-Lymphocyte Technology Tissue Donors Tissues Vaccines Viral Virus antigen-specific T cells comparative cost effective experimental study indexing multiple omics protein expression protein profiling single cell analysis single cell technology single-cell RNA sequencing tool transcriptome

项目摘要

SINGLE CELL CORE: PROJECT SUMMARY The Single Cell Core will provide state-of-the-art experimental technology for single-cell multi-omic profiling. Two major limitations of conventional scRNA-seq for immunophenotyping include the lack of correlation between mRNA levels and surface protein expression and the lack of clonal identity from immune receptor sequence. The Core will provide access to technologies for simultaneous profiling of mRNA, immune receptor sequence, and surface protein expression from individual cells at scale. For experiments with limited cell numbers, we will also provide scPLATE-seq, a fully automated platform for index sorting and scRNA-seq that is cost-effective for low cell numbers. Given the focus on analyzing virus-specific immune responses longitudinally and across tissues, the ability to associate immunophenotypes such as surface protein expression, the transcriptome, and TCR sequence with antigen specificity is also highly desirable. This information will allow us to determine the phenotypic subset and clonal identity of each cell in a virus-specific T cell population, to anchor virus-specific T cells from different individuals, ages, and tissue sites to our reference map, and to investigate their functional capacities with single-cell resolution. The Core will provide scalable technology for associating antigen specificity and T cell receptor sequence with targeted mRNA and surface protein profiles of individual cells (TetTCR- seqHD). We will develop and optimize barcoded tetramer pools for a broad range of viruses and vaccines including CMV, influenza, and SARS-CoV-2 for joint analysis T cell receptor antigen recognition, clonotype, and immunophenotype in individual T cells.

单细胞核心：项目摘要 Single Cell Core 将为单细胞多组学分析提供最先进的实验技术。二传统 scRNA-seq 用于免疫表型分析的主要局限性包括两者之间缺乏相关性 mRNA 水平和表面蛋白表达以及免疫受体序列克隆身份的缺乏。该核心将提供同步分析 mRNA、免疫受体序列、以及单个细胞的表面蛋白大规模表达。对于细胞数量有限的实验，我们将还提供 scPLATE-seq，一个用于索引排序的全自动平台和经济高效的 scRNA-seq 细胞数量低。鉴于重点是纵向和跨领域分析病毒特异性免疫反应组织、关联免疫表型的能力，例如表面蛋白表达、转录组和具有抗原特异性的TCR序列也是非常理想的。这些信息将使我们能够确定病毒特异性 T 细胞群中每个细胞的表型子集和克隆身份，以锚定病毒特异性 T 来自不同个体、年龄和组织部位的细胞到我们的参考图上，并研究它们的功能具有单细胞分辨率的能力。核心将提供可扩展的技术来关联抗原特异性和 T 细胞受体序列以及单个细胞的靶向 mRNA 和表面蛋白谱 (TetTCR- seqHD）。我们将为各种病毒和疫苗开发和优化带条形码的四聚体池包括CMV、流感和SARS-CoV-2，用于联合分析T细胞受体抗原识别、克隆型、和个体 T 细胞的免疫表型。