Production of bovine transgenic embryos for the coagulation factor IX

• Grant number: 08/00102-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Effective date (Start): July 01, 2008
• Effective date (End): June 30, 2010
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Flávio Vieira Meirelles
• Grantee: Paulo Sérgio Monzani
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil

Abstract

Blood coagulation factors are necessary for maintenance of the hemostasis and the deficiency in any factor leads the bleeding disorder. Factors VIII (FVIII) and IX (FIX) deficiencies are responsible for hemophilia A and B, respectively, while factor VII (FVII) deficiency presents similar characteristics of the hemophilia A. These hereditary deficiencies are treated by the administration of such factors for containment bleeding episodes. The supraphysiological doses of FVII can be used to treat uncontrolled bleeding episodes that occur in patients with inhibitors to FVIII and FIX, besides this it is used to preventive case or hemorrhage from traumas and in invasive surgical procedures. Currently, the production of such recombinant factors is carried through cells culture, which presents high cost and limited production for the actual demand. Production of recombinant proteins in milk of transgenic animals has presents the alternative most viable to supply the demand of such factors and to decrease the cost of the same ones. This project proposes amplification of DNA fragments of the promoter and enhancers regions of the gene bovine beta casein. To clone this promoter in pEGFP-N1 vector and to evaluate the capacity of expression of these promoter by production of green fluorescent protein (GFP), in epithelial cells mammary gland. To introduce the promoter with the better expression capacity in pLXIN-FIX vector, with express FIX and was constructed by researchers of Hemocentro (USP - Ribeirão Preto). To transfect epithelial cell mammary gland with the vector constructed and evaluates the FIX expression. Nuclear transfer of transgenic cells mammary gland to bovine enucleated oocytes. Finally, production of transgenic blastocysts for coagulation factor IX.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

BRESSAN, FABIANA F.; SANGALLI, JULIANO R.; PESSA, LAIS V. F.; PIRES, PEDRO R. L.; MEIRELLES, FLAVIO V.. Insights on bovine genetic engineering and cloning. Pesquisa Veterinária Brasileira, v. 33, p. 6-pg., 2013-12-01. (08/57877-3, 11/08376-4, 09/11631-6, 08/00102-0)

BRESSAN, FABIANA F.; SANGALLI, JULIANO R.; PESSA, LAIS V. F.; PIRES, PEDRO R. L.; MEIRELLES, FLAVIO V.. Experiências em clonagem e transgenia em bovinos. Pesquisa Veterinária Brasileira, v. 33, n. 1, p. 113-118, DEC 2013. (08/57877-3, 11/08376-4, 09/11631-6, 08/00102-0)

MONZANI, PAULO S.; ADONA, PAULO R.; OHASHI, OTAVIO M.; MEIRELLES, FLAVIO V.; WHEELER, MATTHEW B.. Transgenic bovine as bioreactors: Challenges and perspectives. BIOENGINEERED, v. 7, n. 3, p. 123-131, 2016. (08/00102-0)