CORE-- VECTOR CORE

载体核心将在专门为病毒载体的高通量生产而配备的设施中设计、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备。载体核心工作人员拥有丰富的经验，包括作为国家服务核心参与NHLBI卓越基因治疗项目，并开发了一套全面的基因转移试剂和方案，并在其使用方面具有专业知识。为了支持该项目，Core将与项目PI密切协调载体生产计划，在相关启动子的控制下，提供多种血清型、报告基因、CFTR基因片段和完整cDNA版本的AAV载体。持续的矢量生产和新矢量构造的持续方法开发是最适合核心的任务

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.