Efficient Cre/loxP site-specific recombination in a HepG2 human liver cell line

Naoya Kobayashi, Hirofumi Noguchi, Karen A. Westerman, Toshihisa Matsumura, Takamasa Watanabe, Toshinori Totsugawa, Toshiyoshi Fujiwara, Philippe Leboulch, Noriaki Tanaka

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12 Citations (Scopus)

Abstract

A worldwide shortage of donor livers is a limiting factor of the clinical application of hepatocyte transplantation (HTX). To resolve this issue, we focused on a reversible immortalization system that allows temporary expansion of primary hepatocyte populations by transfer of an oncogene that can be subsequently excised. As a preliminary test toward this goal, we examined the efficacy of Cre/loxP site-specific recombination in a transformed human liver cell line, HepG2. The present study utilized retroviral transfer of a prototypical immortalizing gene, simian virus 40 large T antigen (SV40Tag), flanked by a pair of loxP recombination targets and adenovirus-mediated Cre/loxP recombination. Here we report that complete elimination of the retroviral transferred oncogene was achieved by site-specific recombination using a replication-deficient recombinant adenovirus vector producing Cre recombinase (Ad-Cre).

Original languageEnglish
Pages (from-to)737-742
Number of pages6
JournalCell Transplantation
Volume9
Issue number5
DOIs
Publication statusPublished - Jan 1 2000

Keywords

  • Cre/loxP site-specific recombination
  • HepG2 cells
  • Reversible immortalization

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cell Biology
  • Transplantation

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    Kobayashi, N., Noguchi, H., Westerman, K. A., Matsumura, T., Watanabe, T., Totsugawa, T., Fujiwara, T., Leboulch, P., & Tanaka, N. (2000). Efficient Cre/loxP site-specific recombination in a HepG2 human liver cell line. Cell Transplantation, 9(5), 737-742. https://doi.org/10.1177/096368970000900525