Determining mechanism of insertational mutagenesis caused by CRISPR/Cas9 genome editing

CRISPR/Cas9基因组编辑引起的插入突变机制的确定

• 批准号: 1803108
• 金额: --
• 依托单位: CARDIFF UNIVERSITY
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1803108
• 关键词: Determining; mechanism; insertational; mutagenesis; caused

Little is currently known about the detailed mechanism of DNA repair initiated by the oligonucleotide-targeted induction of DNA double strand breaks by the CRISPR/Cas9 system. It is currently thought that the doublestrand breaks are repaired by one of the two major double strand break pathways; NHEJ and HR. However, at present, it is not clear what the relative contribution of these two repair pathways in 'on' or 'off'-target CRISPR/Cas9 induced events is. To investigate this we will use novel microarray-based methods developed in Professor Reed's group, for investigating the genomic mechanism of DNA damage induction and its repair in the context of chromatin. This will be combined with the Reed's sophisticated computer algorithms, necessary for the bioinformatics analysis of this data. These technologies have already been developed to investigate the DNA repair mechanisms of genotoxic agents (hence this project will also allow access and validation of these tools for other research applications).

目前对CRISPR/Cas9系统通过寡核苷酸靶向诱导DNA双链断裂启动DNA修复的详细机制知之甚少。目前认为，双链断裂是由两条主要的双链断裂途径之一修复的；NHEJ和HR。然而，目前尚不清楚这两种修复途径在“开”或“关”靶向CRISPR/Cas9诱导事件中的相对贡献是什么。为了研究这一点，我们将使用Reed教授小组开发的基于微阵列的新方法，研究染色质背景下DNA损伤诱导及其修复的基因组机制。这将与里德复杂的计算机算法相结合，这是对这些数据进行生物信息学分析所必需的。这些技术已经被开发用于研究基因毒性药物的DNA修复机制（因此该项目也将允许访问和验证这些工具用于其他研究应用）。