Maranda Vincent, Zhang Yue, Vizeacoumar Frederick S, Freywald Andrew, Vizeacoumar Franco J
Division of Oncology, College of Medicine, University of Saskatchewan, Saskatoon, Canada.
Department of Pathology, College of Medicine, University of Saskatchewan, Saskatoon, Canada.
Methods Mol Biol. 2023;2614:397-409. doi: 10.1007/978-1-0716-2914-7_24.
Large-scale genetic screens are becoming increasingly used as powerful tools to query the genome to identify therapeutic targets in cancer. The advent of the CRISPR technology has revolutionized the effectiveness of these screens and has made it possible to carry out loss-of-function screens to identify cancer-specific genetic interactions. Such loss-of-function screens can be performed in silico, in vitro, and in vivo, depending on the scale of the screen, as well as research questions to be answered. Performing screens in vivo has its challenges but also advantages, providing opportunities to study the tumor microenvironment and cancer immunity. In this chapter, we present a procedural framework and associated notes for conducting in vivo CRISPR knockout screens in cancer models to study cancer biology, anti-tumor immune responses, tumor microenvironment, and predicting treatment responses.
大规模基因筛选正越来越多地被用作强大的工具来查询基因组,以识别癌症中的治疗靶点。CRISPR技术的出现彻底改变了这些筛选的效率,并使得进行功能缺失筛选以识别癌症特异性基因相互作用成为可能。根据筛选的规模以及要回答的研究问题,这种功能缺失筛选可以在计算机上、体外和体内进行。在体内进行筛选有其挑战,但也有优势,为研究肿瘤微环境和癌症免疫提供了机会。在本章中,我们提出了一个程序框架以及相关注释,用于在癌症模型中进行体内CRISPR基因敲除筛选,以研究癌症生物学、抗肿瘤免疫反应、肿瘤微环境以及预测治疗反应。