PRODUCTION OF TRANSGENIC AVIANS USING SPERM-MEDIATED TRANSFECTION

摘要

The present invention relates to novel methods of producing transgenic avians, preferably chickens, wherein the incorporated transgene may be expressed as a constituent protein of the white of a hard-shell egg. The present invention provides sperm-mediated transfer for the introduction to an avian egg of a transgene encoding a heterologous polypeptide. The avian sperm may be irradiated before the transgenic gene is incorporated therein. Transgenic genes may be incorporated into avian sperm by lipofection, electroporation, restriction enzyme mediated integration (REMI) or similar methods. The modified avian sperm may then be delivered to an avian oocyte by microinjection, intracytoplasmic sperm injection (ICSI) or artificial insemination, or by natural coitus after the modified avian sperm are returned to a male bird. Heterologous nucleic acid may be integrated directly into the genomic nucleic acid of the oocyte or after first integrating the heterologous nucleic acid into the nucleic acid of a male germ cell and subsequent delivery of the transgenic male germ cell to an oocyte. Alternatively, the heterologous nucleic acid may be a episome within the sperm, or within the derivative zygote formed by the fusion of the sperm and the recipient oocyte, and may replicate independently of the zygote genome. Co-segregation of the episome with the replicated ooctye genome into all of the daughter cells may be induced by the heterologous nucleic acid having a centromeric body derived from, for example, a chromosome of a chicken.