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研究人员提供载体技术，使其能够在培养和体内高效地将基因转移到哺乳动物细胞，从而促进和促进消化系统疾病的基础和翻译研究。为此，分子生物学和载体核心将：1)作为治疗的教育和咨询资源：DDRCC研究人员，他们可能在病毒衍生的基因转移载体技术方面经验有限，但希望利用这种技术在哺乳动物细胞培养和活体动物模型中高效地功能表达感兴趣的基因序列；2)以最低的成本，提供各种预制的表达标准标记基因的逆转录病毒、慢病毒和腺病毒载体库，用于初步实验，以及表达各种可用载体的库 哺乳动物基因和相应的抑制序列；以及3)以最低的成本设计和生产包含特定目的转基因的定制病毒载体(包括带有或不带有表位标签的野生型和突变型cDNAs、显性负表达结构、反义mRNAs、siRNAs等)。个体化治疗：DDRCC研究人员。通过这种方式，分子生物学和媒介核心将寻求通过向调查人员提供建议、培训、股票媒介试剂、定制媒介来加强CURE：DDRCC计划的活动 生产服务和技术援助。通过这些服务，我们打算促进资助的CURE：DDRCC调查人员的现有研究工作，并为新的调查人员提供获得媒介试剂和技术的机会，这些试剂和技术可能会为未来的拨款申请提供初步数据。此外，通过作为教育和咨询资源，我们寻求向新的研究人员、博士后研究员和研究人员广泛传播最先进的基因转移和表达技术，教他们如何安全地生产和使用病毒载体，并帮助他们准备基因转移。 机构生物安全委员会要求的协议。

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.