Genome-wide CRISPR-Cas9 essentiality screening represents a powerful approach to identify genetic vulnerabilities in cancer cells. Here, we applied this technology and designed a strategy to identify target genes that are synthetic lethal (SL) with von Hippel-Lindau (VHL) tumor suppressor gene. Inactivation of VHL has been frequently found in clear cell renal cell carcinoma. Its SL partners serve as potential drug targets for the development of targeted cancer therapies. We performed parallel genome-wide CRISPR screens in two pairs of isogenic clear cell renal cell carcinoma cell lines that differ only in the VHL status. Comparative analyses of screening results not only confirmed a well-known role for mTOR signaling in renal carcinoma, but also identified DNA damage response and selenocysteine biosynthesis pathways as novel SL targets in VHL-inactivated cancer cells. Follow-up studies provided cellular and mechanistic insights into SL interactions of these pathway genes with the VHL gene. Our CRISPR and RNA-seq datasets provide a rich resource for future investigation of the function of the VHL tumor suppressor protein. Our work demonstrates the efficiency of CRISPR-based synthetic lethality screening in human isogenic cell pairs. Similar strategies could be employed to unveil SL partners with other oncogenic drivers.

中文翻译：

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基因型特异性CRISPR-Cas9筛选发现的VHL合成致死性签名。

全基因组CRISPR-Cas9必需性筛选代表了一种强大的方法来鉴定癌细胞中的遗传脆弱性。在这里，我们应用了这项技术并设计了一种策略，以识别具有von Hippel-Lindau（VHL）肿瘤抑制基因的合成致死（SL）的目标基因。在透明细胞肾细胞癌中经常发现VHL失活。其SL合作伙伴可作为开发靶向癌症疗法的潜在药物靶标。我们在两对仅在VHL状态上不同的等基因透明细胞肾细胞癌细胞系中进行了全基因组CRISPR平行筛选。筛选结果的比较分析不仅证实了mTOR信号在肾癌中的众所周知的作用，而且还确定了DNA损伤反应和硒代半胱氨酸的生物合成途径是VHL灭活的癌细胞中的新型SL靶标。后续研究为这些途径基因与VHL基因的SL相互作用提供了细胞和机制方面的见识。我们的CRISPR和RNA-seq数据集为VHL肿瘤抑制蛋白功能的进一步研究提供了丰富的资源。我们的工作证明了在人类等基因细胞对中基于CRISPR的合成杀伤力筛选的效率。可以采用类似的策略来揭示SL合作伙伴与其他致癌驱动因素。我们的CRISPR和RNA-seq数据集为VHL肿瘤抑制蛋白功能的进一步研究提供了丰富的资源。我们的工作证明了在人类等基因细胞对中基于CRISPR的合成杀伤力筛选的效率。可以采用类似的策略来揭示SL合作伙伴与其他致癌驱动因素。我们的CRISPR和RNA-seq数据集为VHL肿瘤抑制蛋白功能的进一步研究提供了丰富的资源。我们的工作证明了在人类等基因细胞对中基于CRISPR的合成杀伤力筛选的效率。可以采用类似的策略来揭示SL合作伙伴与其他致癌驱动因素。