The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) system has revolutionized the ability to edit the mammalian genome, providing a platform for the correction of pathogenic mutations and further investigation into gene function. CRISPR reagents can be delivered into the cell as DNA, RNA, or pre-formed ribonucleoproteins (RNPs). RNPs offer numerous advantages over other delivery approaches due to their ability to rapidly target genomic sites and quickly degrade thereafter. Here, we review the production steps and delivery methods for Cas9 RNPs. Additionally, we discuss how RNPs enhance genome and epigenome editing efficiencies, reduce off-target editing activity, and minimize cellular toxicity in clinically relevant mammalian cell types. We include details on a broad range of editing approaches, including novel base and prime editing techniques. Finally, we summarize key challenges for the use of RNPs, and propose future perspectives on the field.

成簇的规则间隔短回文重复 (CRISPR)/CRISPR 相关蛋白 (Cas) 系统彻底改变了编辑哺乳动物基因组的能力，为纠正致病突变和进一步研究基因功能提供了平台。CRISPR 试剂可以作为 DNA、RNA 或预先形成的核糖核蛋白 (RNP) 递送到细胞中。与其他递送方法相比，RNP 具有许多优势，因为它们能够快速靶向基因组位点并随后快速降解。在这里，我们回顾了 Cas9 RNPs 的生产步骤和交付方法。此外，我们还讨论了 RNP 如何提高基因组和表观基因组编辑效率，减少脱靶编辑活动，并最大限度地减少临床相关哺乳动物细胞类型中的细胞毒性。我们包括有关各种编辑方法的详细信息，包括新颖的碱基和主要编辑技术。最后，我们总结了使用 RNP 面临的主要挑战，并提出了该领域的未来展望。