CORE--MOLECULAR BIOLOGY AND VECTORS

拟议的CURE: DDRCC分子生物学和载体核心的目标是通过为CURE: DDRCC研究人员提供能够有效地将基因转移到培养和体内哺乳动物细胞的载体技术，促进和促进消化系统疾病的基础和转化研究。为此，分子生物学和载体核心将：1)为CURE: DDRCC研究人员提供教育和咨询资源，这些研究人员可能在病毒衍生的基因转移载体技术方面经验有限，但希望利用这些技术在哺乳动物细胞培养和动物体内模型中有效地表达感兴趣的基因序列；2)以最低的成本，提供表达标准标记基因的各种预制逆转录病毒、慢病毒和腺病毒载体库，用于初步实验，以及表达各种基因的可用载体库

The objective of the proposed CURE: DDRCC Molecular Biology and Vector Core is to promote and facilitate basic and translational research in digestive diseases by providing CURE: DDRCC investigators with access to vector technologies that enable efficient gene transfer to mammalian cells in culture and in vivo. To this end, the Molecular Biology and Vector Core will: 1) serve as an educational and advisory resource for CURE: DDRCC researchers who may have had limited experience with virus-derived gene transfer vector technologies, but who wish to utilize such technologies for efficient functional expression of genetic sequences of interest in mammalian cell culture and in animal models in vivo; 2) at minimal cost, provide various pre-made retroviral, lentiviral, and adenoviral vector stocks expressing standard marker genes to utilize in preliminary experiments, as well as a library of available vectors expressing a variety of mammalian genes and corresponding inhibitory sequences; and 3) at minimal cost, design and produce custom viral vectors that contain a specific transgene of interest (including wild type and mutant cDNAs with or without epitope tags, dominant-negative expression constructs, antisense mRNAs, siRNAs, etc.) for individual CURE: DDRCC researchers. In this way, the Molecular Biology and Vector Core will seek to enhance the activities of the CURE: DDRCC program by providing investigators with advice, training, stock vector reagents, custom vector production services, and technical assistance. Through these services, we intend to facilitate the existing research efforts of funded CURE: DDRCC investigators, and to provide new investigators with access to vector reagents and technologies that may lead to preliminary data for future grant applications. Furthermore, by also serving as an educational and advisory resource, we seek to widely disseminate state-of-the art gene transfer and expression techniques to new investigators, post-doctoral fellows, and research staff, to teach them how to safely produce and use viral vectors, and to assist them in preparing gene transfer protocols required by the Institutional Biosafety Committee.