Enhancing CRISPR-Cas for disease modeling in Xenopus

增强 CRISPR-Cas 在非洲爪蟾疾病模型中的应用

• 批准号: 9900078
• 负责人: Marko E Horb
• 金额: $ 20.75万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9900078
• 关键词: Animal Model; Animals; Architecture; Area; Automobile Driving; Bacteria; Biological Models; Blood; CRISPR/Cas technology; Cell model; Cellular biology; Chromosomes; Clustered Regularly Interspaced Short Palindromic Repeats; Consumption; Development; Disease; Disease model; Exons; Generations; Genes; Genomic DNA; Genomics; Label; Libraries; Limulus; Loligo; Mediating; Messenger RNA; Methods; Modeling; Mutation; Proteins; Refractory; Resources; Specificity; Streptococcus pyogenes; System; Techniques; Temperature; Testing; Time; Variant; Xenopus; Xenopus laevis; case-by-case basis; cold temperature; experience; genome editing; human disease; human model; in vivo; innovation; microbial; non-genetic; novel strategies; precise genome editing

PROJECT SUMMARY The recent development of the CRISPR-Cas system has transformed many non-genetic models of cellular biology, yet the full application of its genome editing capabilities has been largely restricted to warm-blooded species and to genes which are easily amenable to current methods of CRISPR-Cas mediated gene disruption. Several species which live at lower temperatures, such as Xenopus and the marine models Loligo and Limulus, are ideal systems to model human disease, yet current methods of precise genome editing in these species are inadequate. Furthermore, due to their genomic architecture, many highly-conserved disease-causing genes are refractory to current methods of genome editing. We propose to expand the CRISPR-Cas mediated genome editing toolkit in the field of human disease modeling by driving innovation in two areas: 1) the development of a low-temperature CRISPR-Cas system capable of effecting targeted gene insertion in model species which thrive at lower temperatures, and 2) to develop a nascent system of targeted gene disruption by the application of CRISPR-Cas libraries.

项目摘要 CRISPR-CAS系统的最新发展改变了许多细胞的非遗传模型 生物学，但其基因组编辑功能的全部应用主要局限于温水 物种和易于适应CRISPR-CAS介导的基因的方法 破坏。几种生活在较低温度下的物种，例如爪蟾和海洋模型loligo Limulus是建模人类疾病的理想系统，但目前的精确基因组编辑方法 这些物种不足。此外，由于其基因组结构，许多高度保存 引起疾病的基因对当前基因组编辑方法难治性。我们建议扩大 CRISPR-CAS介导的基因组编辑工具包在人类疾病建模领域，通过推动创新 两个领域：1）能够影响靶向基因的低温CRISPR-CAS系统的发展 在较低温度下壮成长的模型物种中的插入，2）开发针对性的新生系统 CRISPR-CAS库的应用中断基因。