Bandyopadhyay Anindya, Kancharla Nagesh, Javalkote Vivek S, Dasgupta Santanu, Brutnell Thomas P
Reliance Industries Ltd., R&D-Synthetic Biology, Navi Mumbai, India.
Chinese Academy of Agricultural Sciences, Biotechnology Research Institute, Beijing China.
Front Plant Sci. 2020 Nov 2;11:584151. doi: 10.3389/fpls.2020.584151. eCollection 2020.
Global population is predicted to approach 10 billion by 2050, an increase of over 2 billion from today. To meet the demands of growing, geographically and socio-economically diversified nations, we need to diversity and expand agricultural production. This expansion of agricultural productivity will need to occur under increasing biotic, and environmental constraints driven by climate change. Clustered regularly interspaced short palindromic repeats-site directed nucleases (CRISPR-SDN) and similar genome editing technologies will likely be key enablers to meet future agricultural needs. While the application of CRISPR-Cas9 mediated genome editing has led the way, the use of CRISPR-Cas12a is also increasing significantly for genome engineering of plants. The popularity of the CRISPR-Cas12a, the type V (class-II) system, is gaining momentum because of its versatility and simplified features. These include the use of a small guide RNA devoid of trans-activating crispr RNA, targeting of T-rich regions of the genome where Cas9 is not suitable for use, RNA processing capability facilitating simpler multiplexing, and its ability to generate double strand breaks (DSB) with staggered ends. Many monocot and dicot species have been successfully edited using this Cas12a system and further research is ongoing to improve its efficiency in plants, including improving the temperature stability of the Cas12a enzyme, identifying new variants of Cas12a or synthetically producing Cas12a with flexible PAM sequences. In this review we provide a comparative survey of CRISPR-Cas12a and Cas9, and provide a perspective on applications of CRISPR-Cas12 in agriculture.
预计到2050年全球人口将接近100亿,比目前增加20多亿。为满足地域和社会经济多样化的国家不断增长的需求,我们需要使农业生产多样化并扩大其规模。农业生产力的这种扩张将需要在气候变化带来的生物和环境限制日益增加的情况下进行。成簇规律间隔短回文重复序列定点核酸酶(CRISPR-SDN)及类似的基因组编辑技术可能是满足未来农业需求的关键推动因素。虽然CRISPR-Cas9介导的基因组编辑的应用已领先一步,但CRISPR-Cas12a在植物基因组工程中的应用也在显著增加。V型(II类)系统CRISPR-Cas12a因其多功能性和简化特性而越来越受欢迎。这些特性包括使用不含反式激活crispr RNA的小向导RNA、靶向基因组中富含T的区域(Cas9不适用于此)、RNA处理能力便于更简单的多重化,以及产生具有交错末端的双链断裂(DSB)的能力。许多单子叶和双子叶物种已使用这种Cas12a系统成功编辑,并且正在进行进一步研究以提高其在植物中的效率,包括提高Cas12a酶的温度稳定性、鉴定Cas12a的新变体或合成产生具有灵活PAM序列的Cas12a。在本综述中,我们对CRISPR-Cas12a和Cas9进行了比较调查,并对CRISPR-Cas12在农业中的应用提出了看法。