CORE--VECTOR FACILITY

将特定的转基因引入细胞(转导)已成为研究和定义细胞和整个动物系统的分子生物学的标准方法。易于转导细胞和组织的重组基因载体是风湿性疾病研究的关键技术。有多种基因转移载体可用于体外和体内的细胞转导，每种类型的基因载体都需要专门的技术来优化基因转移和解决生产问题。这些试剂的构建、表征和制造所需的许多技术都是专门的、昂贵的，如果没有经验的指导很难学习。此外，这些材料的生产需要在专门配置的实验室空间中进行，以符合生物指南。遵守生物安全指南的要求阻止了一些调查人员使用这些有价值的试剂。多个研究人员广泛使用基因载体，支持以核心实验室的形式提供这项技术。自1994年以来，根据多用途关节炎和肌肉骨骼疾病中心(MAMDC)计划，密歇根大学(U-M)风湿病核心中心(RDCC)成员可以获得重组基因载体和相关技术服务。这项目前的提议旨在维持对用于构建、纯化和鉴定含有与关节炎和肌肉骨骼疾病研究相关的基因的重组载体的集中化设施的支持，以用作体外和体内基因转移试剂。载体核心将提供高效和经济高效的病毒和非病毒基因转移技术，包括重组腺病毒、重组MMLV逆转录病毒、重组腺相关病毒、重组慢病毒和表达载体DNA。载体核心公司提供的试剂的质量将有助于将前景看好的基因载体转化为关键的临床前研究和第一阶段人类临床试验。

Introduction of specific transgenes into cells (transduction) has become a standard method used to study and define the molecular biology of both cells and whole animal systems. Recombinant gene vectors that readily transducer cells and tissues represents a critical technology for the study of rheumatic diseases. A variety of gene transfer vectors are available for the transduction of cells both in vitro and in vivo, and each type of gene vector requires specialized technology to optimize gene transfer and address production issues. Many of the techniques required for the construction, characterization and manufacture of these reagents are specialized, expensive and difficult to learn without experienced guidance. In addition, the production of these materials needs to be performed in laboratory space that has been specifically configured in order to comply with biological guidelines. The requirements for compliance with biosafety guidelines has deterred some investigators from pursuing the use of these valuable reagents. The widespread use of gene vectors by multiple investigators supported the provision of this technology in the form of a core laboratory. Recombinant gene vectors and associated technical services have been available to University of Michigan (U-M) Rheumatic Disease Core Center (RDCC) members under the Multipurpose Arthritis and Musculoskeletal Diseases Center (MAMDC) program since 1994. This current proposal is intended to maintain support for a centralized facility for the construction, purification and characterization of recombinant vectors containing genes relevant to the study of arthritis and musculoskeletal diseases for use as in vitro and in vivo gene transfer reagents. The Vector Core will provide efficient and cost effective access to both viral and non-viral gene transfer technologies including; recombinant adenovirus, recombinant MMLV retrovirus, recombinant adeno-associated virus, recombinant lentivirus and expression plasmid DNA. The quality of the reagents provided by the Vector Core will facilitate the translation of promising gene vectors into pivotal preclinical studies and Phase 1 human clinical trials.