Lentiviral Transduction of Neuronal Cells

Here we describe a general method for the construction of a lentivirus vector using a specific example of the construction of a lentivirus containing the luciferase reporter gene under the control of two hypothetical promoters and derived HIV-1 based lentivirus expression vector pLVX-Puro. This method can be used to compare the strength and regulation of different promoters. In this example, the target cells for transduction are human primary fetal astrocytes but the method is applicable to any primary cell culture from the CNS or other tissue and can be used to examine the strength of a particular promoter in different cell types. HIV based lentivirus particles are prepared by transfection of 4 plasmids into 293T cells using the Fugene 6 transfection reagent.