A Xenopus Laevis Research Resource for Immunology

此次更新申请的目标是支持和发展世界上最全面的研究 专门利用两栖动物非洲爪蟾作为多方面的实验资源， 生物医学和免疫学研究的平台，并为整个科学的利益 社区X的兴趣和医学相关性。leevis是由于其免疫的显着相似性， 系统与人类的系统，在所有发展阶段进行实验的可及性，以及 大量遗传和基因组资源的可用性，宝贵的MHC定义的近交系和青蛙克隆 和工具，如淋巴肿瘤细胞系，单克隆抗体，MHC四聚体和电池的验证 免疫相关基因的PCR引物。这些未上市的动物和试剂 需要保存、丰富并提供给科学界。如前所述，两个 提出的主要目标是： (1)保护和促进X。莱维斯免疫生物学研究资源， 管理和分发动物储备和试剂到美国和国外的实验室。我们将保持 并进一步优化我们动物的多样性、质量、生产力和福利。我们将继续提供协助， 培训并告知对使用X感兴趣的科学家、学生和教育工作者。作为研究模型。我们将 经常更新本网站，以提高公众对资源的认识， 我们向科学界传播信息。我们将培养沟通、网络和 与美国和世界上其他非洲爪蟾资源的相互作用。 (2)开发新的方法学和产生新的实验动物和试剂， 主要目标是整合和利用CRISPR/Cas9的最新显着进展和成功- 基于基因组编辑技术产生转基因（Tg）非洲爪蟾。这已被确定为优先事项 由Xenopus社区。具体目标将是：（i）使CRISPR/Cas9系统适应免疫系统， （ii）开发对关键免疫受体具有高度特异性的试剂， （iii）产生和表征荧光Tg报告分子MHC定义的近交系和克隆;和（iv） 继续发展X.用于真实的实时活体显微镜检查。 除了维持一个对非洲爪蟾科学界至关重要的研究平台外， 促进新方法和新技术的发展，可以迅速和广泛地应用于 对组织和器官生理学、免疫学和发育生物学的创新见解。的 这些工具和技术的开发和应用将有助于非洲爪蟾的努力 在美国国立卫生研究院的协助下，社区建立了非洲爪蟾作为生物医学研究的相关模型。

The goal of this renewal application is to support and develop the world's most comprehensive research resource specializing in the use of the amphibian Xenopus laevis as a multi-faceted experimental platform for biomedical and immunological research and for the benefit of the whole scientific community. Interests and medical relevance of X. laevis are due to the remarkable similarity of its immune system with that of human, the accessibility to experimentation at all developmental stages, as well as the availability of large genetic and genomic resources, invaluable MHC-defined inbred strains and clones of frogs and tools such as lymphoid tumor cell lines, monoclonal antibodies, MHC tetramers and batteries of validated PCR primers for immune-relevant genes. These animals and reagents that are not commercially available need to be preserved, enriched, and made available to the scientific community. As in previous proposals, two major main aims are proposed: (1) Preserving and promoting the X. laevis research resource for immunobiology by keeping on managing and distributing animal stocks and reagents to laboratories in the US and abroad. We will maintain and further optimize the diversity, quality, productivity and welfare of our animals. We will continue to assist, train and inform scientists, students and educators interested in using X. laevis as a research model. We will continue to foster the accessibility and public awareness of the resource by frequently updating our web site by which we disseminate information to the scientific community. We will cultivate communication, networking and interactions with other Xenopus resources in US and in the world. (2) Developing new methodologies and generating new experimental animals and reagents, with the major goal of integrating and exploiting the recent remarkable advance and success of the CRISPR/Cas9- based genome editing technology to generate transgenic (Tg) Xenopus. This has been identified as a priority by the Xenopus community. The specific objectives will be: (i) To adapt the CRISPR/Cas9 system for immune gene loss-of-function by transgenesis; (ii) To develop reagents of high specificity for key immune receptors and factors; (iii)To generate and characterize fluorescent Tg reporter MHC-defined inbred lines and clones; and (iv) To continue developing X. laevis tadpoles for real time intravital microscopy. In addition to maintaining a research platform that is crucial for the Xenopus scientific community, this project promotes the development of new approaches and technologies that can be rapidly and broadly applied for innovative insights into tissue and organ physiology, immunology and developmental biology. The development and application of these tools and technologies will contribute to the efforts of the Xenopus community assisted by the NIH to establish Xenopus as a relevant model for biomedical research.