Molecular Biology Core

分子生物学核心

基本信息

批准号：
10628216
负责人：
LIN LIU
金额：
$ 23.94万
依托单位：
OKLAHOMA STATE UNIVERSITY STILLWATER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-05 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10628216
关键词：
2019-nCoV Adenovirus Vector Adopted Award Bacteria Biological Assay Bronchoalveolar Lavage CRISPR interference CRISPR screen CRISPR-mediated transcriptional activation CRISPR/Cas technology Cell Line Cells Clinical Clustered Regularly Interspaced Short Palindromic Repeats Communicable Diseases Consultations DNA Microarray Chip Development Epithelial Cells Gene Silencing Generations Genes Human In Vitro Infectious Diseases Research Institution Knock-out Lentivirus Vector Lung Maintenance Methods Microarray Analysis Molecular Molecular Biology Mutagenesis Nasal Epithelium Non-Viral Vector Oklahoma Organoids Pathway interactions Patients Peripheral Blood Mononuclear Cell Phase RNA Interference Reagent Reporter Research Research Personnel Resources Respiratory Center Respiratory Disease Sampling Services Structure of parenchyma of lung Technical Expertise Techniques Technology Text Time Tissue Model Training Transgenes Transgenic Organisms Virus airway epithelium continuous cell line digital gain of function gene transfer vector genome wide screen genome-wide in vivo loss of function overexpression respiratory respiratory pathogen respiratory virus single-cell RNA sequencing transcriptome sequencing transgene expression

项目摘要

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.

分子生物学核心项目概要利用分子生物学方法来了解呼吸系统疾病的机制由呼吸道病原体引起。分子生物学核心（MBC）的总体目标是提供针对 III 期 CoBRE 和非 CoBRE 的分子生物学集中服务和技术专业知识调查人员。根据第一阶段 CoBRE 奖项，MBC 的成立是为了提供涉及以下方面的服务：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基因检测、实时 PCR 和 DNA 微阵列分析。最先进的基于 CoBRE 研究人员需求的技术和其他服务一直在不断发展自成立以来一直在开发并添加到核心中。在第二阶段，MBC 进行了扩展，包括新的技术，包括 CRISPR/Cas9 技术和批量 RNA 测序，并提供试剂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。该 III 期申请描述了其他尖端技术和有价值的试剂，将向中心研究人员提供包括单细胞 RNA 测序、空间基因分析、基因组广泛的 CRISPR 筛选、人类肺组织模型（肺类器官）和呼吸道临床样本传染病患者。 MBC 的具体目标是为以下方面提供服务和支持：1) 功能性使用 RNAi 和 CRISPR/Cas9、转基因过度表达、诱变和缺失、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 测序和空间基因分析进行广泛基因分析，3) 全基因组 CRISPR 基因激活、敲除和干扰筛选，4) 分子测定，包括报告基因检测、实时 PCR 和液滴数字 PCR，5) 呼吸道和传染病试剂研究包括呼吸道病毒和细菌、报告病毒、细胞系、原代肺细胞、人类呼吸道传染病患者的肺类器官和临床样本，以及6）建议、咨询以及分子生物学各个方面的培训，根据需要开发或采用新方法传播核心资源和技术。