Xenopus models of human disease by targeted genome editing

通过靶向基因组编辑建立人类疾病非洲爪蟾模型

• 批准号: 9257431
• 负责人: Marko E Horb
• 金额: $ 58.31万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-15 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9257431
• 关键词: Adult; Amphibia; Anatomy; Biological Models; Biomedical Research; Brain Diseases; Breeding; Candidate Disease Gene; Cell division; Cells; Cellular biology; Cloning; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Development; Diabetes Mellitus; Diploidy; Disease; Embryo; Fertilization; Fishes; Generations; Genes; Genetic study; Genome; Goals; Grant; Human; Individual; Induced Mutation; Kidney Diseases; Knock-in; Knock-out; Laboratories; Lead; Limb structure; Lung; Methods; Modeling; Modification; Mus; Mutation; Neurobiology; Oocytes; Organogenesis; Paper; Pathway interactions; Physiology; Population; Positioning Attribute; Rana; Reporter; Research; Research Personnel; Resources; Role; Signal Transduction; Situs Inversus; Surveys; System; Technology; Testing; Time; Xenopus; Xenopus laevis; Xenopus oocyte; blastomere structure; body system; congenital heart disorder; cost; design; functional genomics; gain of function; genome editing; human disease; insight; loss of function; meetings; molecular mechanics; mutant; new technology; promoter; public health relevance; repaired; transcription activator-like effector nucleases; xenopus genome

 DESCRIPTION (provided by applicant): Project Summary This proposal supports a project to construct over 100 different Xenopus mutants in key genes related to human disease. The proposed studies will use CRISPR/Cas and TALEN gene editing to create precise models of human disease in the amphibian Xenopus. After a survey of the Xenopus community, there are over 150 different mutants requested to support biomedical research in 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 three main aims to this proposal. First, mutations in key genes will be produced in either Xenopus tropicalis or Xenopus laevis using CRISPR/Cas or TALEN gene editing methods. Second, we propose to develop the use of oocyte host transfer for the generation of mutants using CRISPR/Cas and TALEN methods; this will increase the efficiency with which mutations are induced prior to first embryonic cell division and to allow for the creation of uniform heterozygous mutations by limiting Cas9 activity only in the oocyte. Third, we will optimize knock-in strategies with CRISPR/Cas and/or TALEN. All of these aims will help enhance the utilization of CRISPR/Cas and TALEN gene editing methods in the Xenopus model system.

 描述（由申请人提供）： 项目摘要该提案支持一个项目，以与人类疾病有关的关键基因构建100多种不同的爪蟾突变体。拟议的研究将使用CRISPR/CAS和TALEN基因编辑来在两栖动物中创建人类疾病的精确模型。在对爪蟾社区进行了调查后，要求超过150种不同的突变体支持Xenopus的生物医学研究。当前的研究集中在为整个Xenopus社区开发许多不同疾病的异武模型，每个突变体将与个别研究人员密切协调。该提案有三个主要目标。首先，使用CRISPR/CAS或TALEN基因编辑方法在Tropicalis或Xenopus laevis中将产生关键基因的突变。其次，我们建议使用crispr/cas和talen方法开发使用卵母细胞宿主转移来生成突变体的使用；这将提高在第一次胚胎细胞分裂之前诱导突变的效率，并通过仅在卵母细胞中限制Cas9活性来创建均匀的杂合突变。第三，我们将使用CRISPR/CAS和/或TALEN优化敲门策略。所有这些目标都将有助于增强Xenopus模型系统中CRISPR/CAS和TALEN基因编辑方法的利用。