Molecular Biology Core

分子生物学核心 项目摘要 分子生物学方法被用来理解呼吸系统疾病的潜在机制。 由呼吸道病原体引起。分子生物学核心(MBC)的总体目标是提供 为Cobre和Non-Cobre第三阶段提供集中的分子生物学服务和技术专长 调查人员。在第一阶段Cobre奖下，MBC的成立是为了提供以下服务 分子生物学技术，包括RNAi基因沉默、转基因表达、诱变、基因和 用于基因分析的途径报告分析、实时聚合酶链式反应和DNA微阵列分析。最先进的 基于科布雷调查人员需求的技术和其他服务一直在不断 自核心成立以来一直在开发和增加核心。在第二阶段，MBC扩大到包括新的 技术，包括CRISPR/Cas9技术和批量RNA测序，并为 呼吸道和传染病研究，包括细胞系、原代肺细胞和呼吸道病毒和 细菌。这一第三阶段的申请描述了其他尖端技术和有价值的试剂 将提供给中心研究人员，包括单细胞RNA测序，空间基因图谱，基因组- 广泛的CRISPR筛查、人肺组织模型(肺器官)和呼吸道临床样本 传染病患者。MBC的具体目标是为以下功能提供服务和支持 利用RNAi和CRISPR/Cas9，转基因过表达，突变和缺失，慢病毒， 用于基因转移的腺病毒和非病毒载体，以及稳定的敲除和转基因细胞系，2)基因组- 使用批量RNA测序、单细胞RNA测序和空间基因测序的广泛基因测序，3) 全基因组CRISPR基因激活、敲除和干扰筛选，4)分子分析，包括 报告分析，实时荧光聚合酶链式反应和液滴数字聚合酶链式反应，5)呼吸道和传染病的试剂 研究包括呼吸道病毒和细菌，以及报告病毒，细胞系，原代肺细胞，人类 呼吸道传染病患者的肺器官和临床标本，以及6)建议、咨询 以及关于分子生物学各个方面的培训，根据需要开发或适应新的方法 传播核心资源和技术。

Molecular Biology Core Project Summary Molecular biology approaches are utilized to understand the mechanisms underlying respiratory diseases caused by respiratory pathogens. The overall objective of the Molecular Biology Core (MBC) is to provide centralized services and technical expertise in molecular biology for Phase III CoBRE and non-CoBRE investigators. Under the Phase I CoBRE award, the MBC was established to provide services involving molecular biology technologies, including RNAi gene silencing, transgene expression, mutagenesis, gene and pathway reporter assays, real-time PCR, and DNA microarray analyses for gene profiling. State-of-the-art technologies and other services based on the needs of the CoBRE investigators have been continuously developed and added to the Core since its inception. In Phase II, the MBC was expanded to include new techniques, including CRISPR/Cas9 technology and bulk RNA sequencing, and to offer reagents for respiratory and infectious disease research, including cell lines, primary lung cells, and respiratory viruses and bacteria. This Phase III application describes additional cutting-edge technologies and valuable reagents that will be offered to Center investigators, including single-cell RNA sequencing, spatial gene profiling, genome- wide CRISPR screening, human lung tissue models (lung organoids) and clinical samples from respiratory infectious disease patients. The specific aims of the MBC are to provide services and support for 1) functional studies using RNAi and CRISPR/Cas9, transgene overexpression, mutagenesis and deletion, lentiviral, adenoviral and nonviral vectors for gene transfer, and stable knockout and transgenic cell lines, 2) genome- wide gene profiling using bulk RNA sequencing, single-cell RNA sequencing, and spatial gene profiling, 3) genome-wide CRISPR gene activation, knockout and interference screens, 4) molecular assays including reporter assays, real-time PCR and droplet digital PCR, 5) reagents for respiratory and infectious disease research including respiratory viruses and bacteria, and reporter viruses, cell lines, primary lung cells, human lung organoids and clinical samples from respiratory infectious disease patients, and 6) advice, consultation and training on various aspects of molecular biology, developing or adapting new methods as needed and disseminating core resources and technologies.