Description
• Use this control vector to co-transfect along with lentivirus packaging vectors to make a recombinant control lentivirus.
Lentivirus vector based on the human immunodeficiency virus-1 (HIV-1) has become a promising vector for gene transfer studies. The advantageous feature of lentivirus vector is the ability of gene transfer and integration into dividing and non-dividing cells. The pseudotyped envelope with vesicular stomatitis virus envelope G (VSV-G) protein broadens the target cell range. Lentiviral vectors have been shown to deliver genes to neurons, lymphocytes and macrophages, cell types that previous retrovirus vectors could not be used. Lentiviral vectors have also proven to be effective in transducing brain, liver, muscle, and retina in vivo without toxicity or immune responses. Recently, the lentivirus system is widely used to integrate siRNA efficiently in a wide variety of cell lines and primary cells both in vitro and in vivo.
Lentivirus particles are produced from 293T cells through transient transfection of plasmids that encode for the components of the virion. Due to safety concerns regarding the infectious nature of HIV-1, recent lentiviral packaging systems have separated the viral components into 3 or 4 plasmids. However, these systems still present a small chance of generating replication-competent lentivirus upon recombination. In addition, most commercial lentiviral packaging systems provide plasmids containing the viral structure proteins in a premixed formulation, making it nearly impossible to optimize the ratio of the various plasmids for your particular experiment and host cell.
Lentiviral supernatant can be produced by cotransfecting 293T cells with pLenti-GFP and a lentivirus packaging mix such as MyBioSource's ViraSafe Lentiviral Packaging System. Supernatants can be used directly or purified/concentrated if needed. For long term storage, store supernatant at -80 degree C in aliquots. The resulting GFP control virus can also be used to generate GFP stable cell lines, and stable clones can be selected by green fluorescence sorting