癌症的发病率和死亡率是世界范围内的主要健康问题。除了迄今为止开发的治疗方法外，癌症的不尽人意的治疗效果还没有通过扩大工具箱而得到改善。免疫疗法的出现在实体瘤治疗方面开创了新的时代，但仍然受到限制，需要打破不利影响。同时，通过分子水平的基因分析和操纵可以进一步提高先进技术的发展。尤其是最先进的基因组编辑技术的出现，尤其是集群规则间隔短回文重复序列（CRISPR-Cas9），已经展示了其在癌症免疫疗法方面突破的潜力。在本文中，介绍了CRISPR-Cas9介导的基因组编辑机制和强大的CRISPR工具箱。此外，我们着重审查了CRISPR引起的双链断裂(DSBs)对癌症免疫疗法（敲除或敲入）的影响。最后，我们讨论了基于CRISPR-Cas9的全基因组筛选的靶向识别，强调了空间CRISPR基因组学的潜力，并呈现CRISPR-Cas9在基础研究，转化医学和临床的全面应用和挑战。©2023.作者.

The incidence and mortality of cancer are the major health issue worldwide. Apart from the treatments developed to date, the unsatisfactory therapeutic effects of cancers have not been addressed by broadening the toolbox. The advent of immunotherapy has ushered in a new era in the treatments of solid tumors, but remains limited and requires breaking adverse effects. Meanwhile, the development of advanced technologies can be further boosted by gene analysis and manipulation at the molecular level. The advent of cutting-edge genome editing technology, especially clustered regularly interspaced short palindromic repeats (CRISPR-Cas9), has demonstrated its potential to break the limits of immunotherapy in cancers. In this review, the mechanism of CRISPR-Cas9-mediated genome editing and a powerful CRISPR toolbox are introduced. Furthermore, we focus on reviewing the impact of CRISPR-induced double-strand breaks (DSBs) on cancer immunotherapy (knockout or knockin). Finally, we discuss the CRISPR-Cas9-based genome-wide screening for target identification, emphasis the potential of spatial CRISPR genomics, and present the comprehensive application and challenges in basic research, translational medicine and clinics of CRISPR-Cas9.© 2023. The Author(s).