Application of CRISPR technologies for the efficient development of models of human disease

应用CRISPR技术高效开发人类疾病模型

• 批准号: 2275743
• 金额: --
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2275743
• 关键词: Application; CRISPR; technologies; efficient; development

CRISPR/Cas9 is a programmable site-specific nuclease that can be used to manipulate the sequence of DNA in a precise and efficient manner. Using these "genome editing" approaches, genes can be knocked out of the genome to allow an assessment of their function in both stem cell and animal models. Furthermore, single nucleotide variants which have been found to associate with a particular disease progression or susceptibility can also be introduced into these systems, allowing models of human disease to be interrogated within the laboratory.As well as being an effective genome-editing tool, the CRISPR/Cas9 system, if rendered catalytically inactive, serves as a very effective programmable site-specific DNA binding protein. Using this system, effector domains which can either up- or down-regulate gene expression can be recruited to gene promoters and regulatory regions, allowing tuneable manipulation of gene expression levels - so called CRISPRa (activation) and CRISPRi (inactivation) technology. Since it is becoming clear that much of the genomic variation which influences disease, lies within regulatory regions of the genome, the development of robust systems for manipulating gene expression in models systems will enable insights into how perturbations in gene expression networks underlie changes in disease susceptibility and progression.This research project will focus on efficient methods of implementing and combining CRISPR systems for targeted deletion, inhibition and activation to model disease-associated genomic changes. Working in both stem cells and mouse embryos, the project aims to explore new possibilities for accurate models of human disease, with a particular focus on reducing the animal cost of disease model generation.

CRISPR/Cas9是一种可编程的位点特异性核酸酶，可用于以精确和有效的方式操纵DNA序列。使用这些“基因组编辑”方法，可以将基因从基因组中敲除，以评估它们在干细胞和动物模型中的功能。此外，已经发现与特定疾病进展或易感性相关的单核苷酸变体也可以引入这些系统，从而允许在实验室内研究人类疾病模型。CRISPR/Cas9系统不仅是一种有效的基因组编辑工具，如果使其失去催化活性，还可以作为一种非常有效的可编程位点特异性DNA结合蛋白。使用该系统，可以上调或下调基因表达的效应子结构域可以被募集到基因启动子和调控区，允许基因表达水平的可调操纵-所谓的CRISPRa（激活）和CRISPRi（失活）技术。由于越来越清楚的是，影响疾病的大部分基因组变异都位于基因组的调控区域内，因此开发用于在模型系统中操纵基因表达的强大系统将使人们能够深入了解基因表达网络中的扰动如何导致疾病易感性和进展的变化。该研究项目将专注于实施和组合CRISPR系统的有效方法，抑制和激活以模拟疾病相关的基因组变化。该项目在干细胞和小鼠胚胎中开展工作，旨在探索人类疾病精确模型的新可能性，特别关注降低疾病模型生成的动物成本。