Single Cell Core

单元格核心：项目总结 Single Cell Core将为单细胞多基因组图谱提供最先进的实验技术。二 传统的scRNA-seq用于免疫表型的主要局限性包括缺乏与 MRNA水平和表面蛋白表达，缺乏来自免疫受体序列的克隆性同一性。 核心将提供同时分析信使核糖核酸、免疫受体序列、 以及单个细胞大规模的表面蛋白表达。对于细胞数量有限的实验，我们将 还提供了scPLATE-seq，一个用于索引排序的全自动化平台，以及scRNA-seq，它对以下方面具有成本效益 手机号码少。鉴于重点是纵向和横向分析病毒特异性免疫反应 组织，免疫表型的能力，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细胞群体中每个细胞的表型亚群和克隆性，以锚定病毒特异性T细胞 来自不同个体、年龄和组织部位的细胞到我们的参考图谱，并研究它们的功能 具有单单元分辨率的容量。核心将为关联抗原特异性提供可扩展的技术 和具有单个细胞的靶向mRNA和表面蛋白图谱的T细胞受体序列(TetTCR- Seqhd)。我们将开发和优化条形码四聚体库，用于广泛的病毒和疫苗 包括CMV、流感和SARS-CoV-2联合分析T细胞受体抗原识别、克隆型、 单个T细胞的免疫表型。

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.