Production of transgenic chimeric rabbits and transmission of the transgene through the germline

Abstract

Here we report that improved reproductive technologies combined with an efficient microinjection method and in vitro cultivation medium enabled us to create germ line chimeric rabbits. To follow the fate of the chimeric embryo a blastomere marked with the human blood coagulation factor VIII (hFVIII) transgene was microinjected into a morula stage wild type embryo. The degree of chimerism in different tissues was estimated by real-time PCR and was found to be in the range of 0.1-42%. Among the four chimeric animals, one was identified as a chromosomal intersex and two were germline chimeras.