RESUMEN

Lipofectamine 2000 is a cationic liposome based reagent that provides high transfection efficiency and high levels of transgene expression in a range of mammalian cell types in vitro using a simple protocol. Optimum transfection efficiency and subsequent cell viability depend on a number of experimental variables such as cell density, liposome and DNA concentrations, liposome-DNA complexing time, and the presence or absence of media components such as antibiotics and serum. The importance of these factors in Lipofectamine 2000 mediated transfection will be discussed together with some specific applications: transfection of primary neurons, high throughput transfection, and delivery of small interfering RNAs.

Asunto(s)

Lípidos/química , Neuronas/metabolismo , ARN Interferente Pequeño/genética , Transfección/métodos , Animales , Recuento de Células , Células Cultivadas , Corteza Cerebral/citología , Medios de Cultivo , ADN/genética , Genes Reporteros/genética , Hipocampo/citología , Humanos , Lamina Tipo A/genética , Liposomas , Luciferasas/análisis , Luciferasas/genética , Plásmidos/genética , Interferencia de ARN , Ratas , beta-Galactosidasa/análisis , beta-Galactosidasa/genética