title: Efficient genetic manipulation of 1321N1 astrocytoma cells using lentiviral gene transfer.
abstract: 1321N1 astrocytoma cells are frequently used to analyze stimulus-induced intracellular signaling. These experiments require genetic manipulation of the cells and several chemical and physical methods have been employed in the past. Recently, microporation has been suggested as the best method to transfect 1321N1 astrocytoma cells. Here, we demonstrate that lentiviral gene transfer into 1321N1 cells is highly efficient, cheap and non-toxic. In addition, lentiviral gene transfer efficiently facilitates stable expression of small hairpin RNAs. Finally, lentiviral gene transfer can be used to implant promoter/luciferase reporter genes into the chromatin of the cells, allowing promoter studies using templates that are embedded into the nucleosomal structure of the chromatin.
authors.affiliation: Department of Medical Biochemistry and Molecular Biology, University of Saarland Medical Center, D-66421 Homburg, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Keim
authors.forename: Isabelle
authors.initials: I
authors.lastname: Müller
authors.forename: Gerald
authors.initials: G
authors.lastname: Thiel
mesh_terms: D001254:Astrocytoma / Q000235:genetics*; D001932:Brain Neoplasms / Q000235:genetics*; D045744:Cell Line, Tumor; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D034741:RNA, Small Interfering
chemical_list: D034741:RNA, Small Interfering