Core C: RNA Sequencing Core

核心 C：RNA 测序核心

• 批准号: 9753854
• 负责人: John Christopher Love
• 金额: $ 35万
• 依托单位: YALE UNIVERSITY
• 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/9753854
• 关键词: Autoimmunity; Biopsy; Biotechnology; Blood; Cell Separation; Cells; Complementary DNA; DNA Sequencing Facility; Data; Detection; Disease; Elements; Experimental Autoimmune Encephalomyelitis; Glioma; Health; Human; Immune; Immune response; Immunity; Immunobiology; Immunologics; In Vitro; Institutes; Investigation; Knock-out; Libraries; Love; Malignant Neoplasms; Methods; Molecular Profiling; Mus; Patients; Population; Population Analysis; Preparation; Protocols documentation; Quality Control; RNA Sequences; RNA analysis; Reporting; Reproducibility; Research Personnel; Research Project Grants; Resolution; SLEB2 gene; Sampling; Services; T-Cell Receptor; T-Lymphocyte; Techniques; Technology; Transgenic Mice; base; cell type; comparative; computing resources; differential expression; droplet sequencing; in vivo; interest; monocyte; multiple sclerosis patient; novel; programs; receptor; single cell analysis; single-cell RNA sequencing; tool; transcriptome; transcriptome sequencing

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.

摘要 免疫细胞状态的有效和可重复的检测对于我们理解免疫系统至关重要。 在健康和疾病的背景下，如癌症和自身免疫反应。该服务核心提供 RNA的制备、测序和基本分析的能力和管道。这个实验性的， 计算资源将有助于研究TIGIT、PD 1或PD 2缺陷的EAE小鼠中的免疫力。 从人类活组织检查或多发性硬化症患者血液中分离的细胞的表征， 神经胶质瘤，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 利用了麻省理工学院爱实验室在批量RNA测序方面的专业知识和Shalek实验室在 麻省理工学院与先进的单细胞RNA测序方法。为了实现三个目标， 项目，两个服务将提供：RNA测序的大群体的细胞，和单细胞RNA- 测序（scRNA-Seq）。对于后者，将采用基于平板（96孔或384孔）的测序方法 以及由研究人员Shalek和Love开发的一种名为Seq-Well的新方法， 以类似于报道的Drop-Seq方法的方式平行测序1，000个单细胞。另一 另一方面将是从单细胞回收T细胞受体的能力。这些分析 这一核心的能力将进一步实现该计划的目标， 每个项目都有单细胞分辨率的表征。