尽管新型靶向抗癌药物的开发不断进行，但由于对当前化疗药物产生抗药性，转移性实体瘤的治疗仍然无法达到治愈水平。尽管已经描述了许多药物抗性机制，但对癌细胞通过其他有效的化疗方式逃避的多种方法仍然缺乏一般的理解。在体外分离耐药克隆、定义其耐药机制，并测试这些机制是否在临床药物抗性发挥作用的传统策略耗时且在许多情况下未能提供具有临床意义的信息。在此综述中，我们总结了使用CRISPR技术生成携带定义新型耐药机制的sgRNA的癌细胞文库的方法，包括优点和缺点。描述了使用CRISPR敲除、激活和抑制筛选以及这些方法的组合的现有策略。此外，还描述了专用方法，以识别可能有助于耐药的多个基因，如合成致死性。尽管这些基于CRISPR的方法用于编目癌细胞中药物耐受性基因的应用才刚刚开始，但适当使用它们有望加速对癌症药物耐受性的理解。© 2023年。这是美国政府工作，不受美国版权保护，可能适用于外国版权保护。

Despite the development of new classes of targeted anti-cancer drugs, the curative treatment of metastatic solid tumors remains out of reach owing to the development of resistance to current chemotherapeutics. Although many mechanisms of drug resistance have been described, there is still a general lack of understanding of the many means by which cancer cells elude otherwise effective chemotherapy. The traditional strategy of isolating resistant clones in vitro, defining their mechanism of resistance, and testing to see whether these mechanisms play a role in clinical drug resistance is time-consuming and in many cases falls short of providing clinically relevant information. In this review, we summarize the use of CRISPR technology, including the promise and pitfalls, to generate libraries of cancer cells carrying sgRNAs that define novel mechanisms of resistance. The existing strategies using CRISPR knockout, activation, and inhibition screens, and combinations of these approaches are described. In addition, specialized approaches to identify more than one gene that may be contributing to resistance, as occurs in synthetic lethality, are described. Although these CRISPR-based approaches to cataloguing drug resistance genes in cancer cells are just beginning to be utilized, appropriately used they promise to accelerate understanding of drug resistance in cancer.© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.