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Selection of in vivo retrovirally transduced hepatocytes leads to efficient and predictable mouse liver repopulation
Author(s) -
Guidotti Jacques-Emmanuel,
Mallet Vincent O.,
Mitchell Claudia,
Fabre Monique,
Schoevaert Damien,
Opolon Paule,
Parlier David,
Lambert Martine,
Kahn Axel,
Gilgenkrantz Hélène
Publication year - 2001
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.00-0892fje
Subject(s) - green fluorescent protein , transduction (biophysics) , reporter gene , transgene , biology , plasmid , viral vector , gene , hepatocyte , transfection , in vivo , complementary dna , microbiology and biotechnology , gene expression , in vitro , genetics , biochemistry , recombinant dna
Stable liver gene transfer is generally limited by the low efficiency of commonly used vectors. One way to circumvent this difficulty is to confer a selective advantage on transduced hepatocytes, allowing them to progressively repopulate the liver. We have used a strategy for in vivo selection and controlled amplification of a small percentage of retrovirally engineered hepatocytes. Using a bicistronic retroviral vector encoding a reporter gene and Bcl2, which confers on liver cells a survival advantage against the Fas apoptotic pathway, we demonstrate that 1.5% of initially transduced hepatocytes repopulate up to 85% of the liver after 10 injections of a Fas agonist antibody. Moreover, we show that the kinetics of liver repopulation is highly predictable. This system provides a general means of expanding at will engineered hepatocytes in vivo and offering the possibility to obtain a genetically modified liver expressing a gene of interest in a desired proportion of hepatocytes.