RETROVIRAL INFECTION COUPLED WITH TISSUE-TRANSPLANTATION LIMITS GENE-TRANSFER IN THE CHICKEN-EMBRYO

Gene transfer into early embryos is a powerful methodology for unraveling the molecular bases of developmental processes. One can attempt to minimize widespread effects of an exogenous gene by using tissue- or region-specific promoters in the few instances where they are available. We have developed a method that bypasses the requirement for specific targeting sequences to achieve regionally restricted gene transfer. Intraspecific chimeras have been created by transplantation of restricted portions of a chicken embryo from a donor strain to a host strain. The donor cells are infectable with a recombinant retroviral vector that carries the exogenous gene, whereas the host cells are not. We have demonstrated the feasibility of this approach using a histochemically distinct reporter gene, human placental alkaline phosphatase. The expression of retrovirally transduced alkaline phosphatase was limited to a transplanted hemiprosencephalon (forebrain and eye) in embryonic chickens. This technique can be applied to many other organ systems during avian embryogenesis to test the function(s) of molecules that are normally controlled through spatial and/or temporal regulation, such as many of the growth factor receptors or homeobox-containing proteins.