Molecular Biology Core

分子生物学核心

• 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
• 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

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 项目摘要 利用分子生物学方法来了解呼吸系统疾病的潜在机制 是由呼吸道病原体引起的分子生物学核心（MBC）的总体目标是提供 为III期CoBRE和非CoBRE提供分子生物学方面的集中服务和技术专长 investigators.根据第一阶段的CoBRE合同，MBC的成立是为了提供服务， 分子生物学技术，包括RNAi基因沉默、转基因表达、诱变、基因和 途径报告基因分析、实时PCR和DNA微阵列分析。State-of-the-art 基于CoBRE调查人员需求的技术和其他服务一直在不断地 自成立以来，已开发并添加到核心中。在第二阶段，《禁雷公约》扩大到包括新的 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， 细菌该第三阶段申请描述了额外的尖端技术和有价值的试剂， 将提供给中心的研究人员，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）和来自呼吸系统的临床样品 传染病患者。MBC的具体目标是提供服务和支持1）功能 使用RNAi和CRISPR/Cas9，转基因过表达，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测序和空间基因谱分析的宽基因谱分析，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，以及报告病毒、细胞系、原代肺细胞、人类 来自呼吸道传染病患者的肺类器官和临床样品，以及6）建议、咨询 分子生物学各个方面的培训，根据需要开发或调整新方法， 传播核心资源和技术。