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Abstract: 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.

CRISPR/Cas：一个获得诺贝尔奖的用于基因治疗和作物改良的精准基因组编辑技术

摘要：自从CRISPR/Cas被确认为作为细菌先天性免疫的机制以来，CRISPR/Cas已迅速发展成为功能强大的多功能基因组编辑工具。其中包括开发为单碱基编辑器、引导编辑器、表观遗传编辑器、以及CRISPR干扰（CRISPRi）和CRISPR激活（CRISPRa）的基因调节工具。CRISPR/Cas也可以用于染色质和RNA靶向和成像。在许多生物学领域，尤其是在生物医学和农作物改良方面，CRISPR/Cas的应用已被广泛证明。作为一种新的疾病诊断工具，CRISPR可用于帮助检测和筛查人类和植物疾病，甚至在当前的新型冠状病毒型肺炎（COVID-19）大流行中也得到了应用。CRISPR/Cas也正在作为一种新的基因疗法以治疗包括癌症在内的各种人类疾病，并已帮助开发治疗某些疾病的药物。在农业育种方面，通过CRISPR/Cas精确靶向可以有效地调节关键的分子生物合成，从而用于修饰蛋白质、淀粉、油和其他功能成分来改善作物。除此之外，CRISPR/Cas也被广泛用于调控逆境胁迫相关的基因来显著增强植物对环境胁迫的耐受性和整体作物产量。提高CRISPR/Cas向细胞转化的效率和精度以及限制脱靶活性是该技术广泛应用的主要挑战。

关键词：基因编辑；CRISPR/Cas；新型冠状病毒型肺炎（COVID-19）；癌症；精准育种；作物改良；基因敲入和敲除；基因修复和替代

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