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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Pimentel, J. R. V. De Bem, T. H. C. Monzani, P. S. Sangalli, J. R. Fontes, A. M. Fantinato-Neto, P. Covas, D. T. Meirelles, F. V. Bressan, F. F. Birgel-Junior, E. H. |
| Description | Country affiliation: Brazil Author Affiliation: Monzani PS ( Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, SP, Brasil monzani.paulo@gmail.com.) |
| Abstract | Recombinant coagulation factor IX must be produced in mammalian cells because FIX synthesis involves translational modifications. Human cell culture-based expression of human coagulation factor IX (hFIX) is expensive, and large-scale production capacity is limited. Transgenic animals may greatly increase the yield of therapeutic proteins and reduce costs. In this study, we used a lentiviral system to obtain transgenic cells and somatic cell nuclear transfer (SCNT) to produce transgenic animals. Lentiviral vectors carrying hFIX driven by 3 bovine ß-casein promoters were constructed. Bovine epithelial mammary cells were transduced by lentivirus, selected with blasticidin, plated on extracellular matrix, and induced by lactogenic hormones; promoter activity was evaluated by quantitative PCR. Transcriptional activity of the 5.335-kb promoter was 6-fold higher than the 3.392- and 4.279-kb promoters, which did not significantly differ. Transgenic bovine fibroblasts were transduced with lentivirus carrying the 5.335-kb promoter and used as donor cells for SCNT. Cloned transgenic embryo production yielded development rates of 28.4%, similar to previous reports on cloned non-transgenic embryos. The embryos were transferred to recipient cows (N = 21) and 2 births of cloned transgenic cattle were obtained. These results suggest combination of the lentiviral system and cloning may be a good strategy for production of transgenic cattle. |
| e-ISSN | 16765680 |
| Journal | Genetics and Molecular Research |
| Issue Number | 3 |
| Volume Number | 12 |
| Language | English |
| Publisher | Fundação de Pesquisas Científicas de Ribeirão Preto |
| Publisher Date | 2013-02-28 |
| Publisher Place | Brazil |
| Access Restriction | Open |
| Subject Keyword | Animals, Genetically Modified Breeding Genetics Cloning, Organism Factor Ix Biosynthesis Animals Caseins Chromosome Mapping Dna Fragmentation Embryo, Mammalian Metabolism Epithelial Cells Fibroblasts Cytology Gene Expression Regulation Genetic Vectors Lentivirus Nuclear Transfer Techniques Promoter Regions, Genetic Recombinant Proteins Sequence Analysis, Dna Research Support, Non-u.s. Gov't Discipline Genetics Discipline Molecular Biology Discipline Bioinformatics |
| Content Type | Text |
| Resource Type | Article |
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