Genomics Resources and Infrastucture for the Zebrafish

描述(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，已经产生了数以千计的发育突变，重要的是，大多数导致这些突变表型的基因都有与人类对应的基因，可能会导致相应的异常条件和出生缺陷。虽然这对斑马鱼模型来说是个好兆头，但社区现在有责任利用这个系统提供的巨大潜力来解决与生物医学相关的问题。这一观点最近在提交给美国国立卫生研究院的公共白皮书《斑马鱼作为研究和治疗人类疾病的模式系统》中得到了阐述(见附录)。这次R24竞争更新的首要目标是为个人研究人员提供基因组资源和基础设施，以帮助推动社区进入白皮书要求的更基于功能和目标驱动的生物学。具体地说，我们将：(1)维护公共BAC和大插入筛选资源，以帮助研究人员从无代表性或难以克隆的区域分离基因组克隆；(2)利用经验和系统的方法提高BAC转基因斑马鱼胚胎的效率；(3)为社区创造BAC-Report转基因系；(3)为社区提供具有免疫学意义的基因；以及(4)为本科生实习生和理科教师提供培训环境和研究机会。本申请中提议的核心和资源类活动符合NCRR的R24赠款机制，是本赠款上一个供资周期工作的合乎逻辑的延伸。根据与知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，我们认为对此类BAC公共资源的迫切需求，鉴于我们在BAC建设方面的专业知识，以及现有设备和基础设施已经具备处理和操纵大插入克隆的条件，我们的实验室非常适合履行这一角色。在这项经修订的申请中，我们特别处理了前研究科就目标1和目标4提出的主要关切。 总括而言，我们建议的活动旨在促进斑马鱼研究人员解决与医学有关的研究问题。我们将通过产生对社区广泛使用的基因组资源来做到这一点，这将使研究人员能够更容易地创建人类疾病的动物模型。这些模型很重要，因为它们允许对疾病机制和干预进行基本研究。此外，为了与NCRR的教育使命保持一致，还将通过与当地教育和科学推广计划建立伙伴关系，为本科生和科学教师建立科学推广。

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.