Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory for Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China.
Agronomy Department, University of Florida, Gainesville, FL 32611, USA.
J Zhejiang Univ Sci B. 2021 Apr 15;22(4):253-284. doi: 10.1631/jzus.B2100009.
Since it was first recognized in bacteria and archaea as a mechanism for innate viral immunity in the early 2010s, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) has rapidly been developed into a robust, multifunctional genome editing tool with many uses. Following the discovery of the initial CRISPR/Cas-based system, the technology has been advanced to facilitate a multitude of different functions. These include development as a base editor, prime editor, epigenetic editor, and CRISPR interference (CRISPRi) and CRISPR activator (CRISPRa) gene regulators. It can also be used for chromatin and RNA targeting and imaging. Its applications have proved revolutionary across numerous biological fields, especially in biomedical and agricultural improvement. As a diagnostic tool, CRISPR has been developed to aid the detection and screening of both human and plant diseases, and has even been applied during the current coronavirus disease 2019 (COVID-19) pandemic. CRISPR/Cas is also being trialed as a new form of gene therapy for treating various human diseases, including cancers, and has aided drug development. In terms of agricultural breeding, precise targeting of biological pathways via CRISPR/Cas has been key to regulating molecular biosynthesis and allowing modification of proteins, starch, oil, and other functional components for crop improvement. Adding to this, CRISPR/Cas has been shown capable of significantly enhancing both plant tolerance to environmental stresses and overall crop yield via the targeting of various agronomically important gene regulators. Looking to the future, increasing the efficiency and precision of CRISPR/Cas delivery systems and limiting off-target activity are two major challenges for wider application of the technology. This review provides an in-depth overview of current CRISPR development, including the advantages and disadvantages of the technology, recent applications, and future considerations.
自 21 世纪 10 年代初在细菌和古菌中首次被发现作为先天病毒免疫的机制以来，成簇规律间隔短回文重复序列（CRISPR）/CRISPR 相关蛋白（Cas）迅速发展成为一种具有多种用途的强大多功能基因组编辑工具。在发现最初的基于 CRISPR/Cas 的系统之后，该技术已得到进一步发展，以实现多种不同的功能。这些功能包括开发碱基编辑器、先导编辑工具、表观遗传编辑工具以及 CRISPR 干扰（CRISPRi）和 CRISPR 激活（CRISPRa）基因调控器。它还可用于染色质和 RNA 靶向和成像。它的应用已在众多生物学领域证明具有革命性，特别是在生物医学和农业改良方面。作为一种诊断工具，CRISPR 已被开发用于辅助人类和植物疾病的检测和筛选，甚至在当前的 2019 年冠状病毒病（COVID-19）大流行期间也得到了应用。CRISPR/Cas 也在作为一种新的基因治疗形式进行临床试验，用于治疗包括癌症在内的各种人类疾病，并有助于药物开发。在农业育种方面，通过 CRISPR/Cas 对生物途径进行精确靶向是调节分子生物合成和允许修饰蛋白质、淀粉、油和其他功能成分以改良作物的关键。此外，通过靶向各种具有重要农艺学意义的基因调控器，CRISPR/Cas 已被证明能够显著提高植物对环境胁迫的耐受性和整体作物产量。展望未来，提高 CRISPR/Cas 传递系统的效率和精度并限制脱靶活性是该技术更广泛应用的两个主要挑战。本文深入概述了当前的 CRISPR 发展情况，包括该技术的优缺点、最近的应用以及未来的考虑因素。
