Copyright © 2019 University of Bucharest Rom Biotechnol Lett. 2019; 24(2): 216-228 Printed in Romania. All rights reserved doi: 10.25083/rbl/24.2/216.228 ISSN print: 1224-5984 ISSN online: 2248-3942 *Corresponding author: LILIANA PASLARU, Center of Excellence in Translational Medicine, Fundeni Clinical Institute, 022328, Bucharest, Romania E-mail: lilianaliviapaslaru@yahoo.fr Petruta Alexandru alexpety@yahoo.com, Sanda M. Cretoiu sanda@cretoiu.ro, Simona O. Dima dima.simona@gmail.com, Irinel Popescu prof.irinelpopescu@gmail.com Received for publication, October, 13, 2018 Accepted, December, 27, 2018 Original paper CRISPR/Cas9 gene editing in Huh7 and Hepa RG cell lines LILIANA PASLARU 1*# , PETRUTA ALEXANDRU 2* , SANDA M. CRETOIU 3,* , SIMONA O. DIMA 1 , IRINEL POPESCU 1 1 Center of Excellence in Translational Medicine, Fundeni Clinical Institute, 022328, Bucharest, Romania 2 Department of Molecular Cell Biology, Institute of Biochemistry of the Romanian Academy, 060031, Bucharest, Romania 3 Department of Cell and Molecular Biology and Histology, Carol Davila University of Medicine and Pharmacy, 050474 Bucharest, Romania *authors have equal contributions Abstract Objectives. Mutations and overexpression of β-catenin are associated with many cancers including hepatocellular carcinomas (HCC). Activation of β-catenin signaling may result in resistance to chemotherapeutic agents. The aim of this study was to generate, by CRISPR/Cas9 gene editing technology, a viable cellular model usable to study the detailed mechanisms of HCC and to find target molecules for the development of novel therapeutic drugs. Methods and results. Human Huh7 and HepaRG cells were transfected with CRISPR/CAS β-catenin KO plasmid (h) plus β-catenin HDR plasmid (h). Edited clones were validated by fluorescent microscopy and Western Blot analysis and were further cultured. Additionally, the differential response of parental and KO cells to antitumoral drugs was tested. To our knowledge, this is the first report using CRISPR/Cas9 technology on human HCC cell lines to evaluate the correlation between β-catenin expression and antitumoral drug effects. Conclusion. Our results suggest that β-catenin is possibly involved in chemotherapy resistance, since the Huh7 β-catenin KO cells appeared to be more sensitive compared to Huh7 WT . Keywords CRISPR/cas9, Genome editing, Liver cancer, Wnt/β-catenin signaling pathway, Hepatocellular carcinoma (HCC) To cite this article: PASLARU L, ALEXANDRU P, CRETOIU SM, DIMA SO, POPESCU I. CRISPR/Cas9 gene editing in Huh7 and Hepa RG cell lines. Rom Biotechnol Lett. 2019; 24(2): 216-228. DOI: 10.25083/rbl/24.2/216.228