Efficient liver-directed gene transfer by in situ generation of retroviral vector from adenoviral templates.
To improve liver-directed retroviral-mediated gene transfer, we injected C57/BL10 mice intravenously with three adenoviral vectors encoding retroviral vector genome and structural components: AdGagPol expressing the respective structural genes of Moloney murine leukaemia virus, Ad10A1Env expressing the 10A1 envelope protein of 10A1-MuLV, and AdLEIN, encoding the LEIN retrovirus genome, expressing green fluorescence protein (eGFP) and the neomycin resistance gene.


METHODS
The extent of eGFP expression was determined after 1 and 15 weeks by fluorescence microscopy and FACS analysis. Proviral integration was determined by a novel PCR-based technique.


RESULTS
Hepatocytes infected with all three Ad vectors generated LEIN retrovirus after one week and in situ transduction of neighbouring cells resulted in stable proviral integration associated with eGFP expression ranging from 4.3% to 20.5% in different liver cell populations 15 weeks post-infection.


CONCLUSIONS
Hybrid adeno-retroviral vectors can be efficiently used to improve the efficiency of retroviral-mediated gene transfer to the liver.
Harris
Julian D
JD
SWAN Institute of Biomedical and Life Sciences, School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey, UK.
Roberts
Michael L
ML
Perumal
Dhayaneethie
D
Graham
Ian R
IR
Patterson
Steve
S
Dickson
George
G
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D045744:Cell Line, Tumor; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism / Q000821:virology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008979:Moloney murine leukemia virus / Q000235:genetics*; D014161:Transduction, Genetic; D016662:Virus Integration
D049452:Green Fluorescent Proteins