Transgenic rabbits with genes of human granulocyte colony-stimulating factor and green fluorescent protein

Human granulocyte–colony stimulating factor (GCSF) is one of the pharmacological proteins that can be isolated from the milk of transgenic (TG) animals. The plasmid containing the human GCSF gene under the control of regulatory elements of the bovine β-lactoglobulin gene and the reporter green fluorescent protein (EGFP) gene under the cytomegalovirus (cmv) promoter were obtained. The use of the selected promoters ensures tissue-specific expression of the target protein in the mammary gland of the TG producing animal and a high level of early expression of the reporter protein in eukaryotic cells, which makes it possible to detect TG embryos at the cultivation stage and perform their preimplantation selection. Testing of the gene construct effectiveness was carried out on TG rabbits obtained by microinjection into the male pronucleus of zygotes. It was concluded that GFP is toxic to embryos in the early stages of development due to overexpression of the EGFP gene under a strong cmv promoter. The TG female rabbit (F0) was obtained, in which the level of human GKSF in milk and blood serum was assessed by the ELISA method. Of the 22 baby rabbits obtained from her in four kindling, two were transgenic. Offspring (F1) was obtained from the TG male F0, 56 % of which were males, of which 88 % were TG and did not differ from ordinary rabbits in terms of health. Among females, TG was 10 %, and they died within two weeks after birth.

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