Open AccessGeneration of genetically tailored porcine liver cancer cells by CRISPR/Cas9 editing
Author(s) -
Lobna Elkhadragy,
Meredith M. Regan,
William M. Totura,
Kimia Dasteh Goli,
Shovik Patel,
Kelly García,
Margaret M. Stewart,
Lawrence B. Schook,
Ron C. Gaba,
Kyle M. Schachtschneider
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0119
Subject(s) - crispr , genome editing , cas9 , kras , hepatocellular carcinoma , biology , computational biology , animal model , genetically engineered , genetically modified organism , cancer research , gene , mutation , bioinformatics , genetics , endocrinology
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 TP53 R167H and KRAS G12D 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.