科学家和育种者的长期目标是精确控制基因,以研究其功能并提高作物产量,质量,以及对各种环境压力的耐受性。CRISPR/Cas系统的发现与修改,自然发生的基因编辑工具,开启了研究基因功能和精准作物育种的时代。
在这篇评论中,我们首先介绍了CRISPR/Cas的发现简史,然后介绍了CRISPR/Cas系统的机制和在基因功能研究和作物改良中的应用。目前,CRISPR/Cas基因组编辑已成为作物改良的成熟尖端生物技术工具,已用于作物的许多不同性状。包括病原体抗性,非生物耐受性,植物的发育和形态,甚至次生代谢和纤维发育。最后,指出了与CRISPR/Cas系统相关的主要问题和未来的研究方向。审查的关键科学概念:CRISPR/Cas9系统是一个强大而强大的生物技术工具,用于靶向基因组中的单个DNA和RNA序列。它可以用来定位基因敲入的序列,敲除和替换,以及通过结合特定序列在基因组和表观基因组水平监测和调节基因表达。农杆菌介导的方法仍然是将CRISPR/Cas试剂递送到靶向植物细胞中的主要且有效的方法。然而,其他交付方式,如病毒介导的方法,已经开发并增强了基于CRISPR/Cas9的作物改良的应用潜力。PAM要求提供了CRISPR/Cas9靶向的遗传基因座,也限制了CRISPR/Cas9的应用。发现新的Cas蛋白和修饰当前的Cas酶在基于CRISPR/Cas9的基因组编辑中起着重要作用。开发更好的CRISPR/Cas9系统,包括传递系统和消除脱靶效应的方法,而寻找调控作物生长发育的关键/主基因是基于CRISPR/Cas9的作物改良的两大方向。
It is a long-standing goal of scientists and breeders to precisely control a gene for studying its function as well as improving crop yield, quality, and tolerance to various environmental stresses. The discovery and modification of CRISPR/Cas system, a nature-occurred gene editing tool, opens an era for studying gene function and precision crop breeding.
In this review, we first introduce the brief history of CRISPR/Cas discovery followed the mechanism and application of CRISPR/Cas system on gene function
study and crop improvement. Currently, CRISPR/Cas genome editing has been becoming a mature cutting-edge biotechnological tool for crop improvement that already used in many different traits in crops, including pathogen resistance, abiotic tolerance, plant development and morphology and even secondary metabolism and fiber development. Finally, we point out the major issues associating with CRISPR/Cas system and the future research directions.Key Scientific Concepts of Review: CRISPR/Cas9 system is a robust and powerful biotechnological tool for targeting an individual DNA and RNA sequence in the genome. It can be used to target a sequence for gene knockin, knockout and replacement as well as monitoring and regulating gene expression at the genome and epigenome levels by binding a specific sequence. Agrobacterium-mediated method is still the major and efficient method for delivering CRISPR/Cas regents into targeted plant cells. However, other delivery methods, such as virus-mediated method, have been developed and enhanced the application potentials of CRISPR/Cas9-based crop improvement. PAM requirement offers the CRISPR/Cas9-targted genetic loci and also limits the application of CRISPR/Cas9. Discovering new Cas proteins and modifying current Cas enzymes play an important role in CRISPR/Cas9-based genome editing. Developing a better CRISPR/Cas9 system, including the delivery system and the methods eliminating off-target effects, and finding key/master genes for controlling crop growth and development is two major directions for CRISPR/Cas9-based crop improvement.