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
Vincent W. Keng,
Amy P. Chiu,
Jeffrey C. To,
Xiao-Xiao Li,
Michael A. Linden,
Khalid Amin,
Branden S. Moriarity,
Kosuke Yusa
Affiliations
Vincent W. Keng
The Hong Kong Polytechnic University Shenzhen Research Institute, Shenzhen 518057, China; 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; Corresponding author. 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.
Amy P. Chiu
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
Jeffrey C. To
The Hong Kong Polytechnic University Shenzhen Research Institute, Shenzhen 518057, China; 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
Xiao-Xiao Li
The Hong Kong Polytechnic University Shenzhen Research Institute, Shenzhen 518057, China
Michael A. Linden
Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455, USA
Khalid Amin
Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455, USA
Branden S. Moriarity
Masonic Cancer Center, Department of Pediatrics, and Center for Genome Engineering, University of Minnesota, Minneapolis, MN 55455, USA
Kosuke Yusa
Stem Cell Genetics, Institute for Life and Medical Sciences, Kyoto University, Kyoto, Japan; Wellcome Sanger Institute, Cambridge CB10 1SA, UK; Corresponding author. Stem Cell Genetics, Institute for Life and Medical Sciences, Kyoto University, Kyoto, Japan.
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.
