Oliveros Juan C, Franch Mònica, Tabas-Madrid Daniel, San-León David, Montoliu Lluis, Cubas Pilar, Pazos Florencio
National Centre for Biotechnology (CNB-CSIC), c/ Darwin 3, 28049 Madrid, Spain.
National Centre for Biotechnology (CNB-CSIC), c/ Darwin 3, 28049 Madrid, Spain CIBERER, ISCIII, Madrid, Spain.
Nucleic Acids Res. 2016 Jul 8;44(W1):W267-71. doi: 10.1093/nar/gkw407. Epub 2016 May 10.
The CRISPR/Cas technology is enabling targeted genome editing in multiple organisms with unprecedented accuracy and specificity by using RNA-guided nucleases. A critical point when planning a CRISPR/Cas experiment is the design of the guide RNA (gRNA), which directs the nuclease and associated machinery to the desired genomic location. This gRNA has to fulfil the requirements of the nuclease and lack homology with other genome sites that could lead to off-target effects. Here we introduce the Breaking-Cas system for the design of gRNAs for CRISPR/Cas experiments, including those based in the Cas9 nuclease as well as others recently introduced. The server has unique features not available in other tools, including the possibility of using all eukaryotic genomes available in ENSEMBL (currently around 700), placing variable PAM sequences at 5' or 3' and setting the guide RNA length and the scores per nucleotides. It can be freely accessed at: http://bioinfogp.cnb.csic.es/tools/breakingcas, and the code is available upon request.
CRISPR/Cas技术通过使用RNA引导的核酸酶，能够以前所未有的准确性和特异性在多种生物体中实现靶向基因组编辑。在规划CRISPR/Cas实验时的一个关键点是向导RNA（gRNA）的设计，它将核酸酶及相关机制导向所需的基因组位置。这种gRNA必须满足核酸酶的要求，并且与其他可能导致脱靶效应的基因组位点缺乏同源性。在此，我们介绍用于CRISPR/Cas实验gRNA设计的Breaking-Cas系统，包括基于Cas9核酸酶以及最近引入的其他核酸酶的实验。该服务器具有其他工具所没有的独特功能，包括使用ENSEMBL中所有可用真核基因组（目前约700个）的可能性、在5'或3'端设置可变的原间隔序列临近基序（PAM）序列，以及设置向导RNA长度和每个核苷酸的得分。可通过以下网址免费访问：http://bioinfogp.cnb.csic.es/tools/breakingcas，代码可根据要求提供。
