文章：

利用基于CRISPR的筛选技术识别化疗耐药机制

Use of CRISPR-based screens to identify mechanisms of chemotherapy resistance 

原文发布日期：2023-04-07 

英文摘要：

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. 

摘要翻译： 

尽管新型靶向抗癌药物不断涌现，但由于当前化疗药物耐药性的产生，转移性实体瘤的根治性治疗仍难以实现。虽然已有诸多耐药机制被阐明，但关于癌细胞如何逃逸有效化疗的多种途径仍普遍缺乏认知。传统通过体外分离耐药克隆株、界定其耐药机制并验证这些机制是否参与临床耐药的研究策略，不仅耗时漫长，且在多数情况下难以提供具有临床价值的信息。本综述总结了CRISPR技术的应用——包括其前景与局限——通过构建携带sgRNA的癌细胞文库来揭示新型耐药机制。文中阐述了利用CRISPR基因敲除、激活及抑制筛选的现有策略，以及这些方法的组合应用；同时探讨了识别多个耐药相关基因（如合成致死效应中存在的协同作用）的特异性研究路径。尽管基于CRISPR技术绘制癌细胞耐药基因图谱的研究刚起步，但若能合理应用，必将加速对癌症耐药机制的深入理解。

原文链接：

Use of CRISPR-based screens to identify mechanisms of chemotherapy resistance