Intrauterine gene transfer: gestational stage-specific gene delivery in mice

Intrauterine gene transfer in mice by intraplacental microinjection of recombinant adenoviral or retroviral vectors carrying beta-galactosidase (beta-gal) reporter gene was analyzed in relation to gestational stage, viral titer and promoters. After injections of viral vectors on days 9.5, 11.5 or 14.5 post coitum (p.c.), embryos, fetuses and adult animals were analyzed for beta-gal expression on days 13.5 p.c., 18.5 p.c. and at 2 months of age, respectively. Injection of adenoviral vectors on day 9.5 or day 11.5 p.c. resulted in high beta-gal expression in the heart or liver, respectively. Injection on either day also gave expression in other tissues including vasculature and hindbrain. This temporal pattern of adenoviral transduction correlated with the expression level of integrin beta3 subunit, which is known to be a component involved in adenoviral transduction. Adenovirus-mediated intrauterine gene transfer resulted in persistent beta-gal expression in multiple cell foci in the liver and hearts of 2-month-old adult animals treated in utero, indicating stable integration of the transgene into the host cell genome at a low frequency. Although at low efficiency, injection of retroviral vector on day 9.5 and 11.5 p.c. resulted in beta-gal expression in embryonic liver, while day 9.5 p.c. injection resulted in persistent beta-gal expression in 2-month-old adult heart. This is the first study to show gestational stage-specific gene transfer via intraplacental microinjection of adenoviral vectors.