Online High-throughput Mutagenesis Designer Using Scoring Matrix of Sequence-specific Endonucleases
Online High-throughput Mutagenesis Designer Using Scoring Matrix of Sequence-specific Endonucleases
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DOI：
10.1515/jib-2015-283
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发表时间：
2015-01-01
影响因子：
1.9
通讯作者：
Wang, Hongyu
中科院分区：
文献类型：
--
作者：
Guo, Dayong;Li, Xiaojing;Wang, Hongyu
CRISPR Cas9和其他序列特异性内切酶是支持基因敲除和基因治疗的基本基因组编辑程序。利用这些新技术，需要一个快速、准确的计算等位基因设计者来构建高吞吐量的基因突变管道。创建了一个自动化系统，Cas9在线设计器(COD)，用于筛选Cas9靶点和非靶点，以及提供基因敲除和基因分型策略。在该在线系统的指导下，成功地建立了基因敲除大鼠模型，并进行了基因分型，证实了其预测真实靶点和非靶点的能力。该系统即时设计了72个大鼠细胞色素P450基因突变的基因敲除策略，以证明其高通量效率。此外，该系统还使用了一个非靶标评分矩阵，该矩阵可以应用于除Cas9之外的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、灵活和高通量的计算基因敲除流水线，支持序列特异性内切酶诱导的突变。
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.