Production of transgenic bovine embryos by microinjection of lentiviral vectors or somatic cell nuclear transfer

In these days, the main barriers for production of transgenic animals are the identification of efficient systems of transgenic, such as different mechanisms on finding gene expression. Based on the idea of searching for different ways of developing transgenic animals, the objective of this research was to produce transgenic bovine embryos using the lentiviral vector of microinjection or nuclear transfers of genetically modified somatic cells, which have the finality of developing and quickly selecting animals from transgenic bovine embryos. This research was conducted in two different stages. Stage I consisted on establishing and producing a lentiviral vector which codified GFP protein. Stage II consisted on conducting three different experiments. On experiment I, a microinjection was developed on the lentiviral vector in the perivitelline space of the bovine ovocites before fecundation (T4), obtaining 5,26 percent of blastocites. When compared with the controle (T1) (complex ovoocites with cumulus-ovocites and without manipulation) the percetage was lower (P< 0,05), control without microinjection (T2), or a control with microinjections and Talp medium (T3). On all the microinjected embryos with the lentiviral vectors, a gene expression of GFP (Green Fluourocent Protein) was detected. On experiment II, a microinjection of the lentiveral vector on the perivitelline space of the of the bovine zygote was evaluated six hours after fecundation. The microinjected zygotes with the lentiveral vector (T4), on in vitro cultivars, blastocite rates day seven and day eight, 3,4% and 3,8% respectively. This values were below (P 0.05) in blastocyst rate of embryos treated with trichostatin (T1) (18.1%) when compared with untreated embryos (T2) (6.8%). When the rate was calculated from cleaved zygotes, production was higher (P 0,05) na taxa de blastocistos de embriões tratados com tricostatina (T1) (18,1%) quando comparada com embriões não tratados (T2) (6,8%). Quando a taxa foi calculada a partir de zigotos clivados, a produção foi superior (P