Targeting Usher syndrome with CRISPR mediated base editing

通过 CRISPR 介导的碱基编辑治疗 Usher 综合征

• 批准号: MR/V029924/1
• 负责人: Lauren Major
• 金额: $ 34.3万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FV029924%2F1
• 关键词: Targeting; Usher; syndrome; CRISPR; mediated

Usher syndrome is an hereditary deaf-blindness syndrome inherited in a recessive manner. There are different types of Usher syndrome displaying a range in the severity of symptoms: Type II is the most common and patients demonstrate progressive retinal degeneration occurring after the onset of deafness. Usher syndrome is commonly caused by point mutations in the USH2A gene that result in a defective version of a protein called usherin. It is thought that the usherin protein is involved in long term maintenance of photoreceptors, the cells involved in light detection in the retina.The aim of this DPhil is to correct individual point mutations in the USH2A gene using a technique called base editing. A guide RNA directs the base editing apparatus to a specific section of DNA adjacent to the point mutation so that an enzyme can change the mutated nucleotide to its original form, thus restoring visual acuity. The base editing system will be delivered to patients inside a viral vector via an injection underneath the retina. This method of delivery has already had success in human clinical trials involving other genetic retinal diseases. After the base editing system has been designed, I will begin by testing its efficacy in vitro and will then progress to testing it on a mouse model of Usher syndrome created by the Harwell Institute. Experiments can then be performed to investigate whether any physiological rescue has occurred. It will be important to optimise the base editing system to reduce any unwanted side effects and to maximise its efficiency. The work involved in this DPhil has the potential to make a significant impact on the lives of patients with Usher syndrome. Additionally, if the study is successful it will provide an excellent basis for future base editing experiments directed at reversing harmful genetic mutations both inside and outside the retina.

亚瑟综合症是一种以隐性方式遗传的遗传性聋盲综合症。 Usher 综合征有不同类型，其症状的严重程度各不相同：II 型是最常见的，患者在耳聋发作后表现出进行性视网膜变性。 Usher 综合征通常是由 USH2A 基因的点突变引起的，该突变导致称为 usherin 的蛋白质出现缺陷。据认为，usherin 蛋白参与感光细胞的长期维持，感光细胞参与视网膜中的光检测。该 DPhil 的目的是使用一种称为碱基编辑的技术来纠正 USH2A 基因中的单个点突变。向导RNA将碱基编辑装置引导至与点突变相邻的DNA的特定部分，以便酶可以将突变的核苷酸改变为其原始形式，从而恢复视力。碱基编辑系统将通过视网膜下方注射，通过病毒载体传递给患者。这种输送方法已经在涉及其他遗传性视网膜疾病的人体临床试验中取得了成功。碱基编辑系统设计完成后，我将首先在体外测试其功效，然后在哈韦尔研究所创建的亚瑟综合症小鼠模型上进行测试。然后可以进行实验来调查是否发生了任何生理救援。优化碱基编辑系统以减少任何不需要的副作用并最大限度地提高其效率非常重要。该博士学位所涉及的工作有可能对亚瑟综合症患者的生活产生重大影响。此外，如果这项研究成功，它将为未来旨在逆转视网膜内外有害基因突变的碱基编辑实验提供良好的基础。

项目成果

New CRISPR Tools to Correct Pathogenic Mutations in Usher Syndrome.

• DOI: 10.3390/ijms231911669
• 发表时间: 2022-10-01
• 期刊: International journal of molecular sciences
• 影响因子: 5.6

A Review of CRISPR Tools for Treating Usher Syndrome: Applicability, Safety, Efficiency, and In Vivo Delivery.

• DOI: 10.3390/ijms24087603
• 发表时间: 2023-04-20
• 期刊: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
• 影响因子: 5.6
• 作者: Major, Lauren;McClements, Michelle E.;MacLaren, Robert E.
• 通讯作者: MacLaren, Robert E.