Xenopus Mutant Resource

项目摘要 这笔赠款支持一个项目，以产生和表征非洲爪蟾突变体的关键基因相关的人类 疾病，建立新的非洲爪蟾突变资源，开发新的基因组编辑转基因品系， 建立有效的具体地点一体化。这些研究的长期目标是产生新的模型， 这将为生物医学研究界提供有用的资源。拟议 研究将使用CRISPR-Cas基因编辑在两栖动物中创建人类疾病的精确模型， 爪蟾目前的研究集中在开发许多不同疾病的爪蟾模型， 整个非洲爪蟾群落，每一个突变体都将与个体密切协调发展。 研究人员这项建议有四个主要目的。首先，我们提议与非洲爪蟾合作 研究人员对过去四年中开发的现有突变体（超过116种）进行了表征。的 突变体涵盖了广泛的主题，需要各种各样的专业知识，只能通过以下方式获得 与其他研究人员的互动。第二，我们建议使用CRISPR-Cas产生新的突变体， 研究人员要求。作为非洲爪蟾的国家储备中心，我们拥有经过验证的专业知识， 并维持这些突变体。第三，我们将产生新的转基因品系，用于基因组编辑。 这些新的细胞系将允许Cas9的更多组织特异性表达。第四，我们建议发展 同源性定向修复用于更有效地产生外源DNA的位点特异性整合。所有 这些目标将有助于提高CRISPR-Cas基因编辑方法在非洲爪蟾模型中的应用 系统

Project Summary This grant supports a project to generate and characterize Xenopus mutants in key genes related to human disease, establish a new Xenopus Mutant Resource, develop new transgenic lines for genome editing and establish efficient site-specific integration. The long-term goals of the studies are to generate new models of human disease that will provide useful resources for the biomedical research community. The proposed studies will use CRISPR-Cas gene editing to create precise models of human disease in the amphibian Xenopus. The current studies are focused on developing Xenopus models of many different diseases for the entire Xenopus community, and each mutant will be developed in close coordination with individual researchers. There are four main aims to this proposal. First, we propose to collaborate with Xenopus researchers to characterize existing mutants (over 116) that were developed over the last four years. The mutants cover a wide range of topics that require varied expertise that can only be obtained through interactions with other researchers. Second, we propose to generate new mutants using CRISPR-Cas as requested by researchers. As the national stock center for Xenopus, we have the proven expertise to breed and maintain these mutants. Third, we will generate new transgenic lines that will be used for genome editing. These new lines will allow for more tissue-specific expression of Cas9. Fourth, we propose to develop homology directed repair for more efficient generation of site-specific integration of exogenous DNA. All of these aims will help enhance the utilization of CRISPR-Cas gene editing methods in the Xenopus model system.