Guo Dayong, Li Xiaojing, Zhu Pan, Feng Yanrong, Yang Juan, Zheng Zhihong, Yang Wei, Zhang Enuo, Zhou Shenglai, Wang Hongyu
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J Integr Bioinform. 2015 Mar 1;12(1):35-48. doi: 10.1515/jib-2015-283.
CRISPR Cas9 and other sequence-specific endonucleases are fundamental genome editors supporting gene knockout and gene therapy. A speedy and accurate computational allele designer is required for a high through-put gene mutagenesis pipeline using these new techniques. An automatic system, Cas9 online designer (COD), was created to screen Cas9 targets and off-targets, as well as to provide gene knockout and genotyping strategies. A gene knockout rat model was successfully created and genotyped under the direction of this online system confirming its ability to predict real targets and off-targets. Gene knockout strategies to mutate 72 rat cytochrome P450 genes were designed instantly by the system to demonstrate its high-throughput efficiency. Also, the system used an off-target scoring matrix which can be applied to any sequence-specific genome editing tools besides Cas9. The COD system (http://cas9.wicp.net) has established a speedy, accurate, flexible and high through-put computational gene knockout pipeline supporting the sequence-specific endonuclease induced mutagenesis.
CRISPR Cas9和其他序列特异性核酸内切酶是支持基因敲除和基因治疗的基础基因组编辑工具。对于使用这些新技术的高通量基因诱变流程而言，需要一个快速且准确的计算等位基因设计工具。创建了一个自动系统——Cas9在线设计工具（COD），用于筛选Cas9靶点和脱靶位点，并提供基因敲除和基因分型策略。在这个在线系统的指导下，成功创建了一个基因敲除大鼠模型并进行了基因分型，证实了其预测真实靶点和脱靶位点的能力。该系统即时设计了使72个大鼠细胞色素P450基因发生突变的基因敲除策略，以展示其高通量效率。此外，该系统使用了一个脱靶评分矩阵，除Cas9外，该矩阵还可应用于任何序列特异性基因组编辑工具。COD系统（http://cas9.wicp.net）建立了一个快速、准确、灵活且高通量的计算基因敲除流程，支持序列特异性核酸内切酶诱导的诱变。
