CORE-- VECTOR CORE

核心--矢量核心

• 批准号: 7450949
• 负责人: Stephen Michael Kaminsky
• 金额: $ 23.1万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 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/7450949
• 关键词: Complementary DNA; Core Facility; Cystic Fibrosis Transmembrane Conductance Regulator; Dependovirus; Development; Gene Transfer; Genes; Guanosine Monophosphate; In Vitro; Laboratories; Methods; Modeling; Preparation; Procedures; Production; Protocols documentation; Quality Control; Reagent; Reporter Genes; Schedule; Serotyping; Services; Standards of Weights and Measures; Viral Vector; adeno-associated viral vector; analytical method; design and construction; experience; gene therapy; in vivo; method development; pre-clinical; programs; promoter; vector

The Vector Core will design, construct, produce and characterize adeno-associated virus in a facility equipped specifically for the high through-put production of viral vectors and provide multiple preparations of these vectors to the Program Projects. The Vector Core staffhas extensive experience, including participation as the national service core of the NHLBI Programs in Excellence in Gene Therapy and has developed a comprehensive set of reagents and protocols for gene transfer and expertise in their use. To support this Program Project the Core will closely coordinate the schedule of vector production with the Projects PI's to provide AAV vectors of numerous serotypes, reporter genes, segmental and full cDNA versions of the CFTR gene, under the control of promoters as relevant for the aims. Continuous vector production and on-going methods development for the new vector constructs is a task that best fits the core model in that significant expertise is required and the overall strategy across vector types is similar. The quality of the laboratory grade vectors for in vitro and in vivo applications of 3 of the projects will be verified with analytical methods developed at the Vector Core using standards established for preclinical development in the GMP facility at the Belfer Gene Therapy Core Facility. This quality control mechanism is an integrated part of the Vector Core's program to support the projects and minimize artifactual results. For each of the AAV serotypes the Core uses procedures developed for AAV production that have been optimized for yield, purity and stability of product. Further refinement of methods for each serotype will be provided to assure the quality of vectors prior to use by each of the program projects. The Core estimates it will produce 42 vectors each year for the projects.

Vector Core将在专门用于高通量生产病毒载体的设施中设计、构建、生产和表征腺相关病毒，并为计划项目提供这些载体的多种制剂。Vector Core工作人员拥有丰富的经验，包括作为国家服务核心参与NHLBI卓越基因治疗计划，并开发了一套全面的基因转移试剂和方案及其使用专业知识。为了支持该计划项目，核心将与项目PI密切协调载体生产时间表，以提供多种血清型、报告基因、CFTR基因的片段和完整cDNA版本的腺相关病毒载体，并在与目标相关的启动子的控制下。持续的载体生产和新载体构建的持续方法开发是最适合核心的任务 这是因为需要大量的专业知识，而且各种载体类型的总体战略是相似的。将使用Belfer基因治疗核心机构GMP机构临床前开发的标准，采用Vector Core开发的分析方法，验证其中3个项目的体外和体内应用的实验室级载体的质量。这种质量控制机制是Vector Core项目的一个组成部分，以支持项目并最大限度地减少人为结果。对于每种AAV血清型，核心使用为AAV生产开发的程序，这些程序已针对产物的产率、纯度和稳定性进行了优化。将提供每种血清型方法的进一步改进，以确保每个项目使用前载体的质量。核心估计 每年将为这些项目制作42个载体。