Transferencia de genes in vitro con polímeros catiónicos.
In vitro gene transfer using cationic polymers
Frank Camacho1, Miriela Capo1, Roberto Toledo2, Joel de León3, Arturo Talavera1, Ela M Pérez1.
1Instituto Finlay, Centro de Investigación-Producción de Vacunas. Ave.27 No. 19805. La Lisa. A.P. 16017, C.P. 11600. Ciudad de La Habana, Cuba. E-mail: fcamacho@finlay.edu.cu
2 Centro de Ingeniería Genética y Biotecnología (CIGB). Calle 186 esq. 31, Cubanacán, Playa. A.P. 6162. Ciudad de La Habana, Cuba.
3 Centro de Inmunología Molecular (CIM). Calle 215 esq. 216, Siboney, Playa. Ciudad de La Habana, Cuba.
RESUMEN
La transferencia de genes representa una importante herramienta para el estudio, prevención y tratamiento de enfermedades genéticas y adquiridas así como para estudios relacionados con la función de proteínas de interés. El uso de moléculas catiónicas está ampliamente difundido, ya que su eficiencia en la transfección las convierte en un fuerte rival de otros métodos de transferencia de genes. En este trabajo se transfectaron las líneas celulares COS-7 y BHK-21 con el plásmido psnEGFP que usó los polímeros catiónicos polietilenimina (PEI) y DEAE dextrana. Al medir la eficiencia de transfección se observó que la línea celular BHK-21 deviene en una excelente opción para la transfección con el método de la PEI y que los mejores resultados se obtienen al disolver PEI en NaCl 150 mM. Al centrifugar las placas después de añadidos los complejos ADN-PEI se obtiene cerca del 100% de células transfectadas con bajas concentraciones de ADN.
]]> Palabras claves: Transfección, polietilenimina, PEI, transferencia de genes, polímeros catiónicos.ABSTRACT
Gene transfer represents an important tool for the study, prevention and treatment of genetic and acquired diseases, as well as for protein function studies. Because of their efficiency, the use of cationic molecules has become a strong competitor to other gene transfer methods. In this study, COS-7 and BHK-21 cell lines were transfected with the plasmid psnEGFP using the cationic polymers polyethylene imine (PEI) and DEAE-dextran. When transfection efficiency was measured, it was observed that the cell line BHK-21 is an excellent option for transfection with the PEI method. The best results were obtained when PEI was dissolved in NaCl 150 mM. When plates were centrifuged after addition of the PEI-DNA complexes, nearly 100% of transfected cells with low DNA concentrations were obtained.
Keywords: Transfection, polyethylene imine, PEI, gene transfer, cationic polymers
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