2. Academic Validation
  3. Transposon delivery for CRISPR-based loss-of-function screen in mice identifies NF2 as a cooperating gene involved with the canonical WNT signaling molecular class of hepatocellular carcinoma

Transposon delivery for CRISPR-based loss-of-function screen in mice identifies NF2 as a cooperating gene involved with the canonical WNT signaling molecular class of hepatocellular carcinoma

  • Heliyon. 2023 Jul 28;9(8):e18774. doi: 10.1016/j.heliyon.2023.e18774.
Vincent W Keng 1 2,  Amy P Chiu 2,  Jeffrey C To 1 2,  Xiao-Xiao Li 1,  Michael A Linden 3,  Khalid Amin 3,  Branden S Moriarity 4,  Kosuke Yusa 5 6
Affiliations

Affiliations

  • 1 The Hong Kong Polytechnic University Shenzhen Research Institute, Shenzhen 518057, China.
  • 2 Department of Applied Biology and Chemical Technology, State Key Laboratory of Chemical Biology and Drug Discovery, The Hong Kong Polytechnic University, Kowloon, Hong Kong, China.
  • 3 Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455, USA.
  • 4 Masonic Cancer Center, Department of Pediatrics, and Center for Genome Engineering, University of Minnesota, Minneapolis, MN 55455, USA.
  • 5 Stem Cell Genetics, Institute for Life and Medical Sciences, Kyoto University, Kyoto, Japan.
  • 6 Wellcome Sanger Institute, Cambridge CB10 1SA, UK.
PMID: 37576222 DOI: 10.1016/j.heliyon.2023.e18774
Abstract

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.

Keywords

CRISPR/Cas9; CTNNB1; Hepatocellular carcinoma; NF2; Transposable elements.

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