The clustered regularly interspaced short palindromic repeats (CRISPR)/associated protein 9 (CRISPR/Cas9) genome editing technology is revolutionizing our approach and capability to precisely manipulate the genetic flow of mammalians. The facile programmability of Cas9 protein and guide RNA (gRNA) sequence has recently expanded biomedical application of CRISPR/Cas9 technology from editing mammalian genome to various genetic manipulations. The therapeutic and clinical translation potential of CRISPR/Cas9 genome editing, however, are challenged by its off-target effect and low genome editing efficiency. In this regard, developing new Cas9 variants and conditional control of Cas9/gRNA activity are of great potential for improving genome editing accuracy and on-target efficiency. In this review, we summarize chemical strategies that have been developed recently to engineer the nucleic acid chemistry of gRNA to enhance CRISPR/Cas9 genome editing efficacy, specificity and controllability. This review aims to highlight the endeavor that has been made to solve bottleneck problems in the field of CRISPR/Cas9 and inspire innovative researches to fulfill the gap between bench and bed.

成簇规律间隔的短回文重复序列 (CRISPR)/相关蛋白 9 (CRISPR/Cas9) 基因组编辑技术正在彻底改变我们精确操纵哺乳动物遗传流的方法和能力。Cas9 蛋白和向导 RNA (gRNA) 序列的简便编程性最近将 CRISPR/Cas9 技术的生物医学应用从编辑哺乳动物基因组扩展到各种基因操作。然而，CRISPR/Cas9 基因组编辑的治疗和临床翻译潜力受到其脱靶效应和低基因组编辑效率的挑战。在这方面，开发新的 Cas9 变体和条件控制 Cas9/gRNA 活性对于提高基因组编辑准确性和靶向效率具有巨大潜力。在这篇评论中，我们总结了最近开发的化学策略，以设计 gRNA 的核酸化学，以提高 CRISPR/Cas9 基因组编辑效率、特异性和可控性。这篇综述旨在强调为解决 CRISPR/Cas9 领域的瓶颈问题所做的努力，并激发创新研究以填补工作台和床之间的差距。