肝细胞癌（HCC）存在多种分子亚型，其中涉及许多新型合作致癌基因和肿瘤抑制基因参与其肿瘤发生。已经确定了WNT信号在HCC中的重要性。然而，与这一复杂信号通路相关的复杂遗传机制尚待阐明。重要的是，尽管已经确定了一些合作基因，但仍然存在许多与β-连环蛋白1（CTNNB1）诱导的HCC相关的未知基因。肝细胞癌肿瘤发生需要发生在致癌基因和肿瘤抑制基因中的突变。CRISPR/Cas9系统的出现使研究人员能够高效地靶向两个等位基因。在这项新颖的研究中，Sleeping Beauty转座子系统被用作基因传递系统，将携带gRNA池的表达载体稳定整合到肝细胞基因组中，并过度表达突变的CTNNB1。我们在WNT信号激活背景下鉴定了206个驱动HCC肿瘤发生的候选基因，并在这项原理验证研究中验证了一种具有临床相关性的已知肿瘤抑制基因神经纤维瘤素2（NF2）。© 2023 The Authors.

Various molecular subclasses of hepatocellular carcinoma (HCC) exists, with many novel cooperating oncogenes and tumor suppressor genes involved in its tumorigenesis. The emerging importance of WNT signaling in HCC has been established. However, the intricate genetic mechanisms involved in this complex signaling pathway remains to be elucidated. Importantly, while some cooperating genes have been identified, there are still many unknown genes associated with catenin beta 1 (CTNNB1)-induced HCC. Mutations in both oncogenes and tumor suppressor genes are required for HCC tumorigenesis. The emergence of the CRISPR/Cas9 system has allowed researchers now to target both alleles efficiently. In this novel study, the Sleeping Beauty transposon system was used as a gene delivery system in vivo to stably integrate an expression cassette that carry pools of gRNAs and overexpress a mutant version of CTNNB1 into the hepatocyte genome. We identified 206 candidate genes that drive HCC tumorigenesis in the context of WNT signaling activation and, neurofibromin 2 (NF2) gene, a known tumor suppressor gene with clinical relevance was validated in this proof-of-principle study.© 2023 The Authors.