Lentiviral vectors are able to effectively transduce a broad range of dividing and postmitotic cells. Therefore, they are used for in vitro and in vivo gene delivery, including for the development of gene therapies targeting peripheral organs as well as the central nervous system (CNS).
For production of lentiviral vector particles, we use classical packaging constructs for vector pseudotyping with the vesicular stomatitis virus G protein (VSV-G) envelope. The system is typically based on co-transfection of 3 packaging plasmids, including pCMVDR-8.92, pRSV-Rev, pMD.G, together with the pSIN-WPRE shuttle construct encoding the gene of interest.
The viral particles are produced by transient transfection of HEK293T cells with these four plasmids. High-titer vector stocks compatible with CNS applications can be obtained by ultracentrifugation. The viral titers are determined by p24 antigen measurements, or/and qPCR for infectivity testing.