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

目前对由CRISPR/Cas9系统的DNA双链断裂的阿托伐他汀靶向诱导引发的DNA修复的详细机制知之甚少。目前认为，双链断裂是由两种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“靶上”或“脱靶”CRISPR/Cas9诱导的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，用于研究DNA损伤诱导的基因组机制及其在染色质中的修复。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遗传毒性物质的DNA修复机制（因此，该项目还将允许访问和验证这些工具用于其他研究应用）。

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).