Meaker Grace A, Hair Emma J, Gorochowski Thomas E
School of Biological Sciences, University of Bristol, Bristol BS8 1TQ, UK.
School of Biosciences, Cardiff University, Cardiff CF10 3AT, UK.
Synth Biol (Oxf). 2020 Oct 24;5(1):ysaa021. doi: 10.1093/synbio/ysaa021. eCollection 2020.
The RNA-guided endonuclease system CRISPR-Cas9 has been extensively modified since its discovery, allowing its capabilities to extend far beyond double-stranded cleavage to high fidelity insertions, deletions and single base edits. Such innovations have been possible due to the modular architecture of CRISPR-Cas9 and the robustness of its component parts to modifications and the fusion of new functional elements. Here, we review the broad toolkit of CRISPR-Cas9-based systems now available for diverse genome-editing tasks. We provide an overview of their core molecular structure and mechanism and distil the design principles used to engineer their diverse functionalities. We end by looking beyond the biochemistry and toward the societal and ethical challenges that these CRISPR-Cas9 systems face if their transformative capabilities are to be deployed in a safe and acceptable manner.
RNA引导的核酸内切酶系统CRISPR-Cas9自发现以来已被广泛改造，使其能力远远超越双链切割，扩展到高保真插入、缺失和单碱基编辑。由于CRISPR-Cas9的模块化结构及其组成部分对修饰和新功能元件融合的稳健性，这些创新才得以实现。在这里，我们回顾了目前可用于各种基因组编辑任务的基于CRISPR-Cas9系统的广泛工具包。我们概述了它们的核心分子结构和机制，并提炼出用于设计其多样功能的设计原则。最后，我们将目光从生物化学领域拓展到这些CRISPR-Cas9系统若要以安全且可接受的方式发挥其变革性能力所面临的社会和伦理挑战。
