Core C: RNA Sequencing Core

摘要 对免疫细胞状态的有效和可重复性的检测对于我们理解免疫是至关重要的。 在健康和疾病方面的反应，如癌症和自身免疫。该服务核心提供 用于RNA的准备、测序和基本分析的能力和流水线。这是一项实验性的和 计算资源将促进对TIGIT、PD1或TIGIT缺乏的EAE小鼠的免疫研究 同时，从多发性硬化症患者的活组织或血液中分离出的细胞的特征 以及项目1-3中表达TIGIT和/或PD-1的体外刺激细胞的分析。《核心》 利用麻省理工学院的Love Lab和Shalek Lab在批量RNA测序方面的专业知识 麻省理工学院拥有先进的单细胞RNA测序方法。以促进三个方面的目标 项目，将提供两项服务：大量细胞群体的RNA测序，以及单细胞RNA- 测序(scRNA-Seq)。对于后者，基于平板(96孔或384孔)的测序方法将是 以及由研究人员Shalek和Love开发的名为Seq-Well的新方法，该方法允许 以类似于已报道的Drop-Seq方法的方式平行地对1,000‘S单细胞进行测序。另一个 方面将是从单个细胞中恢复T细胞受体的能力。把这些分析性的 这一核心的能力将通过使深度免疫 每个项目都可以使用单细胞分辨率进行表征。

Abstract Efficient and reproducible detection of immune cell states is paramount to our understanding of the immune responses in the context of health and disease, such as cancer and autoimmunity. This service core provides capabilities and a pipeline for the preparation, sequencing and basic analysis of RNA. This experimental and computational resource will facilitate investigations of immunity in EAE mice that are deficient for TIGIT, PD1 or both, the characterization of cells isolated from human biopsies or blood from patients with multiple sclerosis or glioma, and the analysis of in vitro stimulated cells expressing TIGIT and/or PD-1 in Projects 1-3. The Core leverages the strengths of the Love Lab at MIT with expertise in bulk RNA-sequencing and the Shalek Lab at MIT with advanced approaches for single-cell RNA-sequencing. To facilitate the objectives of the three Projects, two services will be provided: RNA-sequencing of bulk populations of cells, and single-cell RNA- sequencing (scRNA-Seq). For the latter, methods for plate-based (96- or 384-well) sequencing will be available, as well as a novel method developed by Investigators Shalek and Love called Seq-Well that allows sequencing of 1,000’s of single cells in parallel in a manner similar to reported Drop-Seq methods. Another aspect will be the capability to recover the T cell receptors from single cells. Together these analytical capabilities in this Core will further the objectives of the Program by making deep immunological characterization with single-cell resolution available for each of the Projects.