Gebler Christina, Lohoff Tim, Paszkowski-Rogacz Maciej, Mircetic Jovan, Chakraborty Debojyoti, Camgoz Aylin, Hamann Martin V, Theis Mirko, Thiede Christian, Buchholz Frank
Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
Medical Faculty Carl Gustav Carus, Medizinische Klinik und Poliklinik I, Dresden, Germany.
J Natl Cancer Inst. 2016 Aug 30;109(1). doi: 10.1093/jnci/djw183. Print 2017 Jan.
Although whole-genome sequencing has uncovered a large number of mutations that drive tumorigenesis, functional ratification for most mutations remains sparse. Here, we present an approach to test functional relevance of tumor mutations employing CRISPR/Cas9. Combining comprehensive sgRNA design and an efficient reporter assay to nominate efficient and selective sgRNAs, we establish a pipeline to dissect roles of cancer mutations with potential applicability to personalized medicine and future therapeutic use.
尽管全基因组测序已经发现了大量驱动肿瘤发生的突变,但大多数突变的功能验证仍然很少。在这里,我们提出了一种利用CRISPR/Cas9来测试肿瘤突变功能相关性的方法。结合全面的sgRNA设计和高效的报告基因检测来筛选高效且有选择性的sgRNA,我们建立了一个流程来剖析癌症突变的作用,该流程可能适用于个性化医疗和未来的治疗应用。