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

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 µg 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 αs1-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.

Mensurações ultrassonográficas da cisterna da glândula mamária de caprino transgênico

“…Em vista dos atuais trabalhos envolvendo caprinos transgênicos, denotando a fundamental importância da glândula mamária como principal biorreator na produção de proteínas exógenas Recebido em 26 de agosto de 2011 Aceito em 23 de novembro de 2011 *Autor para correspondência (corresponding author) E-mail: darcioitalo@yahoo.com.br (Freitas et al, 2010) Maastricht, Holanda), acoplado a um transdutor convexo de 5MHz. Os exames foram realizados diariamente, do primeiro ao 17º dia de lactação, e, semanalmente, do 18º ao 80º dia.…”

Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

“…1997; Lee et al. 2000; Freitas et al. 2007), as well as having confirmed the feasibility of the technique for large‐scale production (Edmunds et al.…”

Production of recombinant proteins in milk of transgenic and non-transgenic goats

“…Generally, less than 10% of the offspring are transgenic (Baldassarre and Karatzas, 2004). In Brazil, the first goats were born from the microinjected embryos (Freitas et al, 2003) and, later the birth of the first transgenic goat in Latin America was achieved (Freitas et al, 2007). SCNT, combined with molecular biology and cell culture methods, shows a variety of applications.…”

“…Promotors are sequences of a few kilobases located close to the 5' end of the transcribed region of the gene (Houdebine, 2003). In goats, various regulatory sequences of milk specific genes were already isolated and verified in the production of transgenic animals, such as a murine WAP Baldassarre et al, 2004b), goat β-casein (Ko et al, 2000;Parker et al, 2004), bovine β-casein (Lee et al, 1997;Huang et al, 1998) and goat and bovine αs1-casein (Freitas et al, 2007). The expression levels of recombinant proteins (Table 1) vary from one bioreactor to another, which can be related to various factors, such as the expression vector; method utilized for the transfer of genetic material; number of copies and site of insertion of the transgene (Rosen et al, 1996).…”

“…Human G-CSF is widely utilized in different forms of neutropenia, chemotherapeutically induced leukopenia and mobilization of progenitor cells for autologous or allogenic transplants (Welte et al, 1996). Transgenic goats secreting hG-CSF in milk have been already obtained in South Korea (Ko et al, 2000) and Brazil (Freitas et al, 2007;Freitas et al, 2010).…”