Generation of genetically tailored porcine liver cancer cells by CRISPR/Cas9 editing

Lobna Elkhadragy, Maureen R Regan, William M Totura, Kimia Dasteh Goli, Shovik Patel, Kelly Garcia, Matthew Stewart, Lawrence B Schook, Ron C Gaba, Kyle M Schachtschneider

Research output: Contribution to journalArticlepeer-review


Pigs provide a valuable large animal model for several diseases due to their similarity with humans in anatomy, physiology, genetics and drug metabolism. We recently generated a porcine model for TP53R167H and KRASG12D driven hepatocellular carcinoma (HCC) by autologous liver implantation. Here we describe a streamlined approach for developing genetically tailored porcine HCC cells by CRISPR/Cas9 gene editing and isolation of homogenous genetically validated cell clones. The combination of CRISPR/Cas9 editing of HCC cells described herein with the orthotopic HCC model enables development of various porcine HCC models, each with a specific mutational profile. This allows modeling the effect of different driver mutation combinations on tumor progression and in vivo testing of novel targeted therapeutic approaches in a clinically relevant large animal model. METHOD SUMMARY Here we describe a streamlined approach for developing genetically tailored porcine cancer cells. We demonstrate high-efficiency CRISPR/Cas9-mediated gene editing in porcine hepatocellular carcinoma cells and isolation of homogenous gene knockout clones. These genetically tailored cells can be combined with previously described implantation techniques to develop precision porcine models of hepatocellular carcinoma.

Original languageEnglish (US)
Pages (from-to)37-48
Number of pages12
Issue number1
Early online dateNov 23 2020
StatePublished - Dec 2020


  • CRISPR/Cas9
  • Gene editing
  • Gene knockout
  • Hepatocellular carcinoma
  • Large animal model
  • Liver cancer
  • Porcine cells

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology
  • Biotechnology

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