Blin Kai, Pedersen Lasse Ebdrup, Weber Tilmann, Lee Sang Yup
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Alle 6, 2970 Hørsholm, Denmark.
Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Republic of Korea.
Synth Syst Biotechnol. 2016 Feb 12;1(2):118-121. doi: 10.1016/j.synbio.2016.01.003. eCollection 2016 Jun.
CRISPR/Cas9-based genome editing has been one of the major achievements of molecular biology, allowing the targeted engineering of a wide range of genomes. The system originally evolved in prokaryotes as an adaptive immune system against bacteriophage infections. It now sees widespread application in genome engineering workflows, especially using the endonuclease Cas9. To utilize Cas9, so-called single guide RNAs (sgRNAs) need to be designed for each target gene. While there are many tools available to design sgRNAs for the popular model organisms, only few tools that allow designing sgRNAs for non-model organisms exist. Here, we present CRISPy-web (http://crispy.secondarymetabolites.org/), an easy to use web tool based on CRISPy to design sgRNAs for any user-provided microbial genome. CRISPy-web allows researchers to interactively select a region of their genome of interest to scan for possible sgRNAs. After checks for potential off-target matches, the resulting sgRNA sequences are displayed graphically and can be exported to text files. All steps and information are accessible from a web browser without the requirement to install and use command line scripts.
基于CRISPR/Cas9的基因组编辑技术一直是分子生物学的重大成就之一，它能对多种基因组进行靶向工程改造。该系统最初在原核生物中作为一种抵御噬菌体感染的适应性免疫系统进化而来。如今，它在基因组工程工作流程中得到了广泛应用，尤其是使用核酸内切酶Cas9。为了利用Cas9，需要为每个靶基因设计所谓的单向导RNA（sgRNA）。虽然有许多工具可用于为常见模式生物设计sgRNA，但能够为非模式生物设计sgRNA的工具却很少。在此，我们展示了CRISPy-web（http://crispy.secondarymetabolites.org/），这是一个基于CRISPy的易于使用的网络工具，用于为任何用户提供的微生物基因组设计sgRNA。CRISPy-web允许研究人员交互式地选择他们感兴趣的基因组区域，以扫描可能的sgRNA。在检查潜在的脱靶匹配后，生成的sgRNA序列将以图形方式显示，并可导出到文本文件中。所有步骤和信息都可以通过网络浏览器访问，无需安装和使用命令行脚本。
