Lentiviruses are a very reliable class of viral vectors wildly used in gene therapy. In this chapter, we described a general method for the construction of lentiviral delivery system by using a derived HIV-1 based lentivirus expression vector pKLV-Puro containing a monomeric blue fluorescent protein mammalian codon-optimized (TagBFP). HIV-1 based lentivirus particles are prepared by transfection of four plasmids into 293 T cells using the Fugene 6 transfection reagent. In this case, 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.
Keywords: Lentiviral vectors; Primary human fetal astrocytes; Transduction.