Abstract
Objective: To produce bovine transgenic embryos by microinjection of a lentiviral vector that carries the eGFP gene as a marker in zygotes six hours after fertilization. Methods: 834 oocytes were matured and subjected to one of four treatments designed as follows: CC: Control: IVF with Cumulus-oocyte with (COCs), cultivated in CR2 medium supplemented with 10% FBS and incubated at 38.5°C in atmosphere of 95% humidity and 5% CO₂.CCM: Control culture medium: fertilized in vitro for six hours, cultured in medium SOF aluminum in pouches under the same conditions of CC. MC: Microinjection control: Fertilized under the same treatment conditions CCM. After six hours they were microinjected with TALP medium and cultured in sachets with the same conditions of CCM treatment. ML: Microinjected with the lentivirus: Fertilized in the same conditions of the CCM treatment. After six hours they were microinjected with the lentiviral vector carrying the eGFP transgene and cultured in sachets with the same treatment conditions CCM and MC. Findings: The cleavage rate found in CC was higher (p 0.05) in them, but yes, with ML (p < 0.05). On average, 76.4% of the zygotes obtained in ML expressed the green fluorescent protein. Application/Improvements: The cult ure conditions used were suitable for CC, CCM and MC, microinjection with lentiviral vector has some influence on embryo development, it succeeded in obtaining transgenic zygotes. Keywords: Green Fluorescent Protein (eGFP), Genetic Modification, Micromanipulation of Zygotes
Highlights
Modified Organisms (GMOs) or transgenic organisms are organisms that have DNA sequences from another species inserted in their genome[1, 2]
For CC, the cleavage rate obtained was higher (p < 0.05) than that observed in treatments CCM, MC and microinjected with lentivirus (ML)
This fact can be explained by the manipulation to which the possible zygotes of the treatments CCM, MC and ML were subjected, since, six hours after the start of the IVF, these were removed from the drop of fertilization and subjected to the complete removal of cumulus cells
Summary
Modified Organisms (GMOs) or transgenic organisms are organisms that have DNA sequences from another species inserted in their genome[1, 2]. Transgenic Animals (TA) have many applications, such as in vivo study of gene function in organogenesis, development and aging, as experimental models for the knowledge of mechanisms involved in the development of diseases and in animals Farm. This technique helps in breeding programs with the multiplication of animals with desirable characteristics of economic interest[5,6,7]. These techniques vary in efficiency[1]
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