Cell Model & Assay Core

细胞模型

• 批准号: 7288651
• 负责人: KEVIN L KIRK
• 金额: $ 19.72万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7288651
• 关键词: Binding; Biological Assay; Cell Line; Cell model; Cells; Cost Savings; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Development; Dyes; Epithelial Cells; Equipment; Fostering; Funding; Human; Human Cell Line; Human Resources; In Vitro; Individual; Interdisciplinary Study; Investigational Therapies; Laboratories; Laboratory Research; Lung; Measurement; Measures; Membrane; Modeling; Modification; Monitor; Mus; Mutation; Numbers; Patch-Clamp Techniques; Pathogenesis; Peptides; Pharmaceutical Preparations; Protocols documentation; Reagent; Regulation; Research; Research Personnel; Resources; Scientist; Series; Solutions; Techniques; Testing; United States National Institutes of Health; Validation; Zinc Fingers; cellular transduction; endonuclease; experience; inhibitor/antagonist; innovation; metabolomics; mutant; novel; patch clamp; ranpirnase; research study; tool

Cystic fibrosis investigators within the P30 Center require access to expertise and specialized equipment for their studies of CFTR expression and function. For both established CF laboratories and those new to CF research, a need has been identified for developing new cell lines expressing mutant and wild type CFTR, and for dedicated equipment and biophysical protocols necessary to assess CFTR function. In particular, we propose two specific aims: Specific Aim 1. Cellular models expressing wild type and mutant CFTR will be developed and provided by Core A. These will include primary cells from murine and human lungs, novel lentiviral transduced cell lines, and innovative CF models derived by zinc finger endonuclease modification of the CFTR locus in vitro. Specific Aim 2. Assistance, equipment and expertise necessary to perform functional assays of CFTR in the above cell models (Aim 1) will include fluorescent (halide sensitive) dye protocols, Ussing chamber assays and biophysical techniques (patch clamp) necessary to investigate CFTR activity and regulation. The Core will aid in the development/validation of new CF cell models; provide primary murine airway epithelial cells encoding specific CFTR mutations; and assist investigators with their experiments to test the effects of AF508 on CFTR protein stability and channel function. Collaborative studies involving metabolomic consequences of mutations in CFTR, discovery of peptides from the first cytosolic loop of CFTR that specifically block NBD1 TMD1 binding and numerous other NIH funded projects will also be assisted by the Core. Core A will foster interdisciplinary research by providing valuable new cell models and assays of CFTR expression and function to investigators less familiar with the requisite techniques, and contribute to innovative studies of CF pathogenesis and experimental therapy.

P30中心内的囊性纤维化调查人员需要使用专业知识和专业设备 他们对CFTR表达和功能的研究。对于已建立的CF实验室和CF新手的实验室 研究，已经确定了开发表达突变体和野生型CFTR的新细胞系的需求， 以及用于评估CFTR功能所需的专用设备和生物物理方案。特别是我们 提出了两个具体目标： 特定目的1。表达野生型和突变体CFTR的细胞模型将由 核心A.这些将包括来自鼠和人肺的原代细胞，新型慢病毒转导的细胞系， 以及通过锌指内切酶修饰CFTR基因座的创新CF模型。 特定目标2。执行CFTR功能测定所需的帮助，设备和专业知识 上述细胞模型（AIM 1）将包括荧光（卤化物敏感）染料方案，使用室 研究CFTR活性和调节所需的测定和生物物理技术（贴片夹）。 核心将有助于开发/验证新的CF细胞模型；提供主要鼠气道 编码特定CFTR突变的上皮细胞；并协助研究人员进行实验以测试 AF508对CFTR蛋白稳定性和通道功能的影响。涉及的协作研究 CFTR突变的代谢组后果，从第一个胞质环发现的肽 专门阻止NBD1 TMD1绑定和许多其他NIH资助项目的CFTR也将是 由核心协助。 核心A将通过提供有价值的新细胞模型和CFTR测定来促进跨学科研究 对调查人员的表达和功能不太熟悉所需的技术，并有助于 CF发病机理和实验疗法的创新研究。