EF1α-RFP-Puro Lentivirus

Lentivirus is currently the most commonly used viral gene introduction system and the main gene introduction platform used in cell and gene therapy research, especially CAR-T therapy. Lentivirus is a single-stranded RNA virus, which is generally developed based on human immunodeficiency virus (HIV-1). It can be used for gene introduction in most mammalian cells, including non-dividing cells such as hematopoietic stem cells and neurons, which are difficult to achieve using retroviruses. The target gene introduced using recombinant lentivirus can also be integrated into the genome of the host cell, which can achieve long-term and stable gene expression.

The wild-type lentiviral genome is about 9.7 kb (including two LTRs). Artificially constructing a genome longer than this will result in unstable viral particles and a significant decrease in viral titer. For lentiviral expression vectors, most of the viral genome has been replaced by other useful sequences, such as selection markers or fluorescent protein genes, but there is still enough space for cloning the target gene. Because each lentiviral expression vector contains different useful sequences, the available space for cloning transgenes is also different. Optimized lentiviral expression vectors, in addition to containing HIV-1 LTR and lentiviral packaging signal (Ψ), also contain some specific elements (cPPT/CTS, RRE, WPRE), the presence of which helps to improve transgene expression, viral titer and overall lentiviral vector function.