Xie Xianrong, Li Fuquan, Tan Xiyu, Zeng Dongchang, Liu Weizhi, Zeng Wanyong, Zhu Qinlong, Liu Yao-Guang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, China.
Comput Struct Biotechnol J. 2022 Jul 29;20:4009-4014. doi: 10.1016/j.csbj.2022.07.046. eCollection 2022.
CRISPR-dependent base editors enable direct nucleotide conversion without the introduction of double-strand DNA break or donor DNA template, thus expanding the CRISPR toolbox for genetic manipulation. However, designing guide RNAs (gRNAs) for base editors to enable gene correction or inactivation is more complicated than using the CRISPR system for gene disruption. Here, we present a user-friendly web tool named BEtarget dedicated to the design of gRNA for base editing. It is currently supported by 46 plant reference genomes and 5 genomes of non-plant model organisms. BEtarget supports the design of gRNAs with different types of protospacer adjacent motifs (PAM) and integrates various functions, including automatic identification of open reading frame, prediction of potential off-target sites, annotation of codon change, and assessment of gRNA quality. Moreover, the program provides an interactive interface for users to selectively display information about the desired target sites. In brief, we have developed a flexible and versatile web-based tool to simplify complications associated with the design of base editing technology. BEtarget is freely accessible at https://skl.scau.edu.cn/betarget/.
依赖CRISPR的碱基编辑器能够实现直接的核苷酸转换,而无需引入双链DNA断裂或供体DNA模板,从而扩展了用于基因操作的CRISPR工具箱。然而,为碱基编辑器设计引导RNA(gRNA)以实现基因校正或失活比使用CRISPR系统进行基因破坏更为复杂。在此,我们展示了一个名为BEtarget的用户友好型网络工具,专门用于碱基编辑的gRNA设计。目前它得到46个植物参考基因组和5个非植物模式生物基因组的支持。BEtarget支持设计具有不同类型原间隔序列临近基序(PAM)的gRNA,并整合了多种功能,包括开放阅读框的自动识别、潜在脱靶位点的预测、密码子变化的注释以及gRNA质量评估。此外,该程序为用户提供了一个交互式界面,以便选择性地显示所需靶位点的信息。简而言之,我们开发了一个灵活通用的基于网络的工具,以简化与碱基编辑技术设计相关的复杂性。可通过https://skl.scau.edu.cn/betarget/免费访问BEtarget。