Center for Experimental and Molecular Medicine, Cancer Center Amsterdam and Amsterdam Gastroenterology Endocrinology Metabolism, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, Amsterdam 1105AZ, The Netherlands; Oncode Institute, Meibergdreef 9, Amsterdam 1105AZ, The Netherlands.
Center for Experimental and Molecular Medicine, Cancer Center Amsterdam and Amsterdam Gastroenterology Endocrinology Metabolism, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, Amsterdam 1105AZ, The Netherlands; Oncode Institute, Meibergdreef 9, Amsterdam 1105AZ, The Netherlands.
STAR Protoc. 2023 Dec 15;4(4):102588. doi: 10.1016/j.xpro.2023.102588. Epub 2023 Sep 28.
Long non-coding RNAs (lncRNAs) are emerging as key regulators in the initiation, growth, and progression of cancer. High-throughput CRISPR-based techniques systematically assess the function of genes or regulatory elements present in the human genome. Here, we present a protocol for identifying essential lncRNAs in cancer using CRISPRi-based dropout screens. We describe steps to select target sites, design guide RNAs, and generate CRISPRi cell lines. We then detail the execution and analysis of CRISPRi-based dropout screens.
长链非编码 RNA（lncRNAs）作为关键调控因子，在癌症的发生、生长和进展中发挥着重要作用。高通量基于 CRISPR 的技术系统地评估了人类基因组中存在的基因或调控元件的功能。在这里，我们提出了一种使用基于 CRISPRi 的敲低筛选来鉴定癌症中必需 lncRNA 的方案。我们描述了选择靶位点、设计向导 RNA 和生成 CRISPRi 细胞系的步骤。然后，我们详细介绍了基于 CRISPRi 的敲低筛选的执行和分析。
