Single Cell Core

单电池芯：项目总结 单细胞核心将为单细胞多组学分析提供最先进的实验技术。两 用于免疫表型分析的常规scRNA-seq的主要局限性包括缺乏与免疫表型之间的相关性。 mRNA水平和表面蛋白表达以及缺乏来自免疫受体序列的克隆同一性。 核心将提供同时分析mRNA、免疫受体序列、 和单个细胞的表面蛋白表达。对于细胞数量有限的实验，我们将 还提供scPLATE-seq，一个用于索引排序和scRNA-seq的全自动化平台， 细胞数量少。鉴于重点是纵向和横向分析病毒特异性免疫反应， 组织，将免疫表型如表面蛋白表达，转录组， 具有抗原特异性的TCR序列也是高度期望的。这些信息将使我们能够确定 在病毒特异性T细胞群体中每个细胞的表型亚群和克隆身份，以锚病毒特异性T 从不同的个体，年龄和组织部位的细胞到我们的参考图谱，并研究其功能 单细胞分辨率的容量。核心将提供可扩展的技术， 和T细胞受体序列与单个细胞的靶向mRNA和表面蛋白质谱（TetTCR-1）。 seqHD）。我们将为广泛的病毒和疫苗开发和优化条形码四聚体池 包括CMV、流感病毒和SARS-CoV-2进行联合分析，T细胞受体抗原识别，克隆分型， 和免疫表型。

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.