Expression of active human blood clotting factor VIII in mammary gland of transgenic rabbits.

Human clotting factor VIII is probably the largest protein to be expressed to date in the mammary gland of a transgenic animal, and it requires extensive posttranslational modification to achieve full biological activity. The mammary gland specific construct mWAP-hFVIII-MT-I was injected into the pronuclei of rabbit zygotes, and three transgenic offspring were obtained. Founder 385 showed germ-line transmission of a single integrated copy, and a homozygous line was established from this animal. The rhFVIII was transcribed and translated exclusively in the mammary gland. The activity of rhFVIII in the rabbit milk ranged from 5 to 8% of that found in normal human plasma. Results indicate the suitability of the transgenic rabbit mammary gland for rhFVIII production.

[1]  E. Gócza,et al.  Production of transgenic chimeric rabbits and transmission of the transgene through the germline , 2004, Molecular reproduction and development.

[2]  S. Wu,et al.  Temporal and spatial expression of biologically active human factor VIII in the milk of transgenic mice driven by mammary-specific bovine α-lactalbumin regulation sequences , 2002, Transgenic Research.

[3]  J. Smeitink,et al.  Enzyme therapy for Pompe disease with recombinant human α-glucosidase from rabbit milk , 2001, Journal of Inherited Metabolic Disease.

[4]  K. Garber.  rFactor VIII deficit questioned , 2000, Nature Biotechnology.

[5]  A. Dove.  Milking the genome for profit , 2000, Nature Biotechnology.

[6]  T. Raju,et al.  Species-specific variation in glycosylation of IgG: evidence for the species-specific sialylation and branch-specific galactosylation and importance for engineering recombinant glycoprotein therapeutics. , 2000, Glycobiology.

[7]  J. Grisham.  New rules for Bt corn , 2000, Nature Biotechnology.

[8]  J. de la Fuente,et al.  Transgenic rabbits for the production of biologically-active recombinant proteins in the milk. , 1999, Genetic analysis : biomolecular engineering.

[9]  H. Niemann,et al.  Expression of human blood clotting factor VIII in the mammary gland of transgenic sheep , 1999, Transgenic Research.

[10]  G. Brem,et al.  Species-specific alternative splicing of transgenic RNA in the mammary glands of pigs, rabbits, and mice. , 1999, Biochemical and biophysical research communications.

[11]  F. Gwazdauskas,et al.  Transgenic pigs produce functional human factor VIII in milk , 1997, Nature Biotechnology.

[12]  D. Kerr,et al.  Transgenic dairy cattle: genetic engineering on a large scale. , 1997, Journal of dairy science.

[13]  A. Subramanian,et al.  Rate Limitations in Posttranslational Processing by the Mammary Gland of Transgenic Animals a , 1996, Annals of the New York Academy of Sciences.

[14]  R. Offringa,et al.  A quick method to estimate the T-DNA copy number in transgenic plants at an early stage after transformation, using inverse PCR , 1991, Plant Molecular Biology.

[15]  R. Kaufman,et al.  Structure-Function Relationships of Factor VIII Elucidated through Recombinant DNA Technology , 1989, Thrombosis and Haemostasis.

[16]  G. Brem,et al.  Generation and Application of Transgenic Rabbits , 1998 .

[17]  C. Holt,et al.  The composition of milk. , 1983 .