Alok Anshu, Sandhya Dulam, Jogam Phanikanth, Rodrigues Vandasue, Bhati Kaushal K, Sharma Himanshu, Kumar Jitendra
University Institute of Engineering and Technology, Panjab University, Chandigarh, India.
Department of Biotechnology, Kakatiya University, Warangal, India.
Front Plant Sci. 2020 Mar 31;11:264. doi: 10.3389/fpls.2020.00264. eCollection 2020.
Cpf1, an endonuclease of the class 2 CRISPR family, fills the gaps that were previously faced in the world of genome engineering tools, which include the TALEN, ZFN, and CRISPR/Cas9. Other simultaneously discovered nucleases were not able to carry out re-engineering at the same region due to the loss of a target site after first-time engineering. Cpf1 acts as a dual nuclease, functioning as an endoribonuclease to process crRNA and endodeoxyribonuclease to cleave target sequences and generate double-stranded breaks. Additionally, Cpf1 allows for multiplexed genome editing, as a single crRNA array transcript can target multiple loci in the genome. The CRISPR/Cpf1 system enables gene deletion, insertion, base editing, and locus tagging in monocot as well as in dicot plants with fewer off-target effects. This tool has been efficiently demonstrated into tobacco, rice, soybean, wheat, etc. This review covers the development and applications of Cpf1 mediated genome editing technology in plants.
Cpf1是2类CRISPR家族的一种核酸酶，它填补了此前基因组工程工具领域所面临的空白，这些工具包括转录激活样效应因子核酸酶（TALEN）、锌指核酸酶（ZFN）和CRISPR/Cas9。其他同时发现的核酸酶由于首次工程操作后靶位点的丢失，无法在同一区域进行重新工程操作。Cpf1作为一种双功能核酸酶，作为核糖核酸内切酶加工crRNA，作为脱氧核糖核酸内切酶切割靶序列并产生双链断裂。此外，Cpf1允许进行多重基因组编辑，因为单个crRNA阵列转录本可以靶向基因组中的多个位点。CRISPR/Cpf1系统能够在单子叶植物和双子叶植物中进行基因缺失、插入、碱基编辑和基因座标记，且脱靶效应较少。该工具已在烟草、水稻、大豆、小麦等植物中得到有效验证。本综述涵盖了Cpf1介导的植物基因组编辑技术的发展和应用。
