Core C: RNA Sequencing Core

核心 C：RNA 测序核心

基本信息

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
项目类别：
财政年份：
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

项目摘要

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.

抽象的免疫细胞态的有效且可重复的检测对于我们对免疫的理解至关重要在癌症和自身免疫等健康和疾病的背景下的反应。此服务核心提供 RNA的制备，测序和基本分析的功能和管道。这个实验和计算资源将有助于调查EAE小鼠的免疫力，而Tigit，PD1或这两者都是从人类活检中分离出的细胞的表征或来自多发性硬化症患者的血液或血液的表征胶质瘤，以及在项目1-3中表达Tigit和/或PD-1的体外刺激细胞的分析。核心利用MIT的爱实验室的优势，并具有散装RNA的专业知识和Shalek Lab的专业知识 MIT采用高级方法进行单细胞RNA测序。促进三个目标的目标将提供项目，两项服务：大量细胞的RNA序列，单细胞RNA- 测序（SCRNA-SEQ）。对于以后的，基于板的方法（96-或384孔）测序将是可用，以及调查人员Shalek和Love称为Seq-Well的新方法，允许以类似于报道的滴序方法类似的方式将1,000个单元的测序平行地进行。其他方面将是从单个细胞中恢复T细胞受体的能力。这些分析在一起该核心的能力将通过深入免疫学来进一步进一步每个项目都可以使用单细胞分辨率进行表征。