Production of transgenic goat (Capra hircus) with human Granulocyte Colony Stimulating Factor (hG-CSF) gene in Brazil

Vicente J.F Freitas,Dárcio I.A Teixeira,Karlliely C Almeida,Edilson S Lopes-Jr,João B Cajazeiras,Lyudmila E Andreeva,Suely R.G Avelar,Antonio C.C Carvalho,Luciene P.B Dias,Irina A Serova,Francisco C Sousa,Oleg L Serov,Luciana M Melo,Guennadi A Dvoryanchikov,Maria L.L Andrade,Alexsandra F Pereira,Raylene R Moura

doi:10.1590/s0001-37652007000400003

Abstract

In order to produce transgenic goats with hG-CSF, a total of 24 adult Saanen and 48 adult undefined breed goats were used as donors and recipients, respectively. Donors were estrus-synchronized with vaginal sponges and superovulated by a treatment with 200 mg FSH given twice daily in decreasing doses over 3 days starting 48 h before sponge removal. Ovulation was induced by injecting 100 microg GnRH 36 h after sponge removal. The recipients also received an estrus synchronization treatment. Donors were mated with fertile Saanen bucks and, approximately 72 h after sponge removal, zygotes were recovered surgically by flushing oviducts. The recovered zygotes were briefly centrifuged to a reliable visualization of the pronuclei. The DNA construct containing hG-CSF gene flanked by goat and bovine alphas1-casein sequences was injected into pronuclei of 129 zygotes. The microinjected embryos (3-6 per female) were transferred to 27 recipients. Ten recipients became pregnant and 12 kids were born. One transgenic male founder was identified in the group of kids. This is the first report of a birth of a transgenic goat in Latin America.

Highlights

Production of valuable proteins of pharmaceutical interest in the milk of transgenic farm animals has become an attractive alternative to microbial and animal cell bioreactors
In order to produce transgenic goats with human Granulocyte Colony Stimulating Factor (hG-CSF), a total of 24 adult Saanen and 48 adult undefined breed goats were used as donors and recipients, respectively
The DNA construct containing hG-CSF gene flanked by goat and bovine αs1-casein sequences was injected into pronuclei of 129 zygotes

Summary

Introduction

Production of valuable proteins of pharmaceutical interest in the milk of transgenic farm animals has become an attractive alternative to microbial and animal cell bioreactors. The human Granulocyte Colony Stimulating Factor (hG-CSF) is a hematopoietic growth factor that stimulates the proliferation and the differentiation commitment of neutrophil precursor cells, and enhances some of the functional properties of mature neutrophils (Morstyn and Burgess 1988). Following its production as a recombinant human protein (Souza et al 1986), hG-CSF has been the most widely used hematopoietic growth factor due to its proven efficacy against different forms of neutropenia and chemotherapy induced leucopenia. Ko et al (2000) have developed a transgenic female goat harboring goat-casein promoter/hG-CSF fusion gene by microinjection into fertilized one-cell goat zygotes

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