Genomics Resources and Infrastucture for the Zebrafish

描述（由申请人提供）：斑马鱼，Danio rerio，已成为研究发育的重要脊椎动物模型系统。通过ENU和插入突变筛选已经产生了数千种发育突变，重要的是，导致这些突变表型的大多数基因在人类中都有对应的基因，可能会导致相应的异常状况和出生缺陷。虽然这对斑马鱼模型来说是个好兆头，但现在社区有责任利用该系统提供的巨大潜力来解决生物医学相关的问题。最近，美国国立卫生研究院的一份公共白皮书《斑马鱼作为研究和治疗人类疾病的模型系统》（见附录）明确表达了这种观点。本次R24竞争性更新的首要目标是为个体研究人员提供基因组资源和基础设施，以帮助社区按照白皮书的要求进入更基于功能和目标驱动的生物学领域。具体而言，我们将：(1)维护公共BAC和大插入筛选资源，以帮助研究人员从未代表或难以克隆的区域分离基因组克隆；(2)通过实证和系统的方法提高斑马鱼胚胎BAC转基因的效果；(3)为基因群体生成bac报告基因转基因系；具有免疫学重要性的；(4)为本科生实习生和理科教师提供培训环境和科研机会。本申请中提出的核心和资源类活动与NCRR的R24拨款机制保持一致，是该拨款前一个资助周期所开展工作的合理延伸。根据与著名斑马鱼研究人员的讨论以及对当前斑马鱼基因组计划数据库的查询，我们认为迫切需要这样的BAC公共资源，我们的实验室非常适合履行这一角色，因为我们在BAC建设方面的专业知识，以及现有的设备和基础设施已经适合处理和操纵大插入克隆。在修订后的申请中，我们特别解决了先前研究部分针对目标1和目标4提出的主要问题。

DESCRIPTION (provided by applicant): The zebrafish, Danio rerio, has emerged as an important vertebrate model system for the study of development. Thousands of developmental mutants have been generated via ENU and insertional mutagenesis screens, and, importantly, the majority of the genes responsible for these mutant phenotypes have human counterparts that may give rise to corresponding aberrant conditions and birth defects. While this augurs well for the zebrafish model, the onus is now on the community to use the tremendous potential offered in this system to address problems of biomedical relevance. This sentiment was recently articulated in a communal White Paper to NIH, "Zebrafish as a model system to study and cure human disease" (see Appendix). The overriding objective of this R24 competitive renewal is to provide genomic resources and infrastructure to individual investigators in order to help move the community into more function-based and goal-driven biology as mandated by the White Paper. Specifically, we will: (1) maintain a communal BAC and large-insert screening resource to aid investigators with isolation of genomic clones from unrepresented or difficult-to-clone regions; (2) improve the efficacy of BAC transgenesis of zebrafish embryos using an empirical and systematic approach; (3) generate BAC-reporter transgenic lines for the community for genes; of immunological importance; and (4) provide a training environment and research opportunities for undergraduate interns and science teachers. The core- and resource-type activities proposed in this application are in keeping with the R24 grant mechanism at NCRR and are logical extensions of the work performed during the previous funding cycle of this grant. Based on the discussion with prominent zebrafish researchers as well as queries of the current zebrafish genome project databases, we believe there is a pressing need for such BAC communal resources and that our laboratory is well-suited to fulfill this role given our expertise in BAC construction and the fact that existing equipment and infrastructure are already it place for handling and manipulating large-insert clones. In this revised application, we have specifically addressed major concerns raised by the previous Study Section with respect to Aims 1 and 4. In summary, our proposed activities are aimed at facilitating zebrafish researchers to address medically relevant research problems. We will do this by generating genomic resources that will be of wide use to the community and which will allow investigators to more easily create animal models of human disease. These models are important as they allow basic investigation into disease mechanism and intervention. In addition in keeping with the educational mission of NCRR, science outreach will be established for undergraduates and science teachers through partnerships with local educational and science outreach programs.