Transfection of Gametes: A Method to Generate Transgenic Animals
Transfección de Gametos: Un Método para Generar Animales Transgénicos
Int. j. morphol; 23 (3), 2005
Publication year: 2005
The production of transgenic animals (TA) using transfected spermatozoa or eggs is commented. Different methods have been employed to introduce transgenes into the gametes of several vertebrates and invertebrates. Methods for the transfection of gametes have employed naked DNA, viral vectors, DNA/Liposome complexes, electroporation or high velocity microprojectiles. Spermatozoa and oocytes or eggs have showed a good transfection efficience (80% in some cases), and microscopical observations demonstrated that the transgenes appeared localized in the nucleus. Gametes have shown to be naturally protected against the entrance of foreign genes because some semino plasma or plasma membrane proteins block the entrance of foreign genes in spermatozoa. In the female this blockage is undertaken by the egg covers (the zona pellucida in mammals and the perivitelline coat in mollusks). In several cases the production of TA has been described after using the transfected gametes for in vitro fertilization or inseminations. Sometimes, larger percentages of TA were observed (85% in salmon). Nevertheless, these TA were mainly chimeric for the transgene and they were not capable to establish transgenic lines. It seems probable that TA produced by gamete transfections are different from those originated by the conventional microinjection procedures. Furthermore, gametes would develop some kind of mechanisms that modify the integration/expression of transgenes, or that block the integration of transgenes in the germinal line.
Se comentan algunos aspectos sobre la transfección de gametos y su empleo para la producción de animales transgénicos (AT). Para la transfección de gametos han sido empleados ADN desnudo, vectores virales, complejos ADN-Liposomas, electroporación, o microproyectiles de alta velocidad. En general, se han obtenido altos porcentajes de transfección (hasta del 80% en espermatozoides) y se ha observado que los transgenes se localizan en el interior del núcleo.