DNA Transfer to Neuronal Primary Cultures Using TransFast™ Transfection Reagent

by Fabrice Ango and Laurent Fagni (E-mail: fagni@bacchus.montp.inserm.fr) UPR CNRS 9023, CCIPE 141 rue de la Cardonille, 34094 Montpellier, France

TransFast ™ Transfection Reagent (Cat.#E2431) is a recent addition to Promega’s line of transfection reagents. We describe an optimized method that is based on the ability of this cationic polymer to transfer cDNA into neuronal primary cultures. Using TransFast™ Reagent, we transfected cultured cerebellar granule cells, as well as other cultured mammalian neuronal cell types, with green fluorescent protein (GFP) cDNA, at a transfection efficiency of at least 15%. This reagent has proved successful in the simultaneous cotransfection of up to three different cDNAs. The procedure is virtually non-neurotoxic as evidenced by normal in vitro development and bioelectrical properties of the transfected neurons. Because this method of transfection is quite easy and can be successfully applied to mammalian neuronal preparations, it provides the singular advantage for the rapid study of a series of eukaryotic proteins under appropriate physiological conditions.

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