Efficient Cre/loxP site-specific recombination in a HepG2 human liver cell line.
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).
Kobayashi
N
N
First Department of Surgery, Okayama University Medical School, Japan. ntanaka@med.okayama-u.ac.jp
Noguchi
H
H
Westerman
K A
KA
Matsumura
T
T
Watanabe
T
T
Totsugawa
T
T
Fujiwara
T
T
Leboulch
P
P
Tanaka
N
N
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D002455:Cell Division; D002461:Cell Line, Transformed; D003673:Defective Viruses / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015774:Ganciclovir / Q000494:pharmacology; D015870:Gene Expression; D005839:Gentamicins / Q000494:pharmacology; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000821:virology*; D006801:Humans; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D016513:Mice, SCID; D009374:Neoplasms, Experimental / Q000473:pathology; D012333:RNA, Messenger / Q000096:biosynthesis; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D014764:Viral Proteins
D000952:Antigens, Polyomavirus Transforming; D005839:Gentamicins; D012333:RNA, Messenger; D014764:Viral Proteins; C010680:antibiotic G 418; C045073:Cre recombinase; D019426:Integrases; D015774:Ganciclovir