EF1α-tdTOMATO-Puro Lentivirus

The packaging system of lentiviral vectors generally consists of two parts, namely packaging components and vector components. The packaging component is constructed by removing the cis-acting sequences required for packaging, reverse transcription and integration from the HIV-1 genome, and can provide the proteins necessary for the production of viral particles in trans. The vector component is complementary to the packaging component, that is, it contains the cis-acting sequences required for packaging, reverse transcription and integration, and has a multiple cloning site under the control of a heterologous promoter and the target gene inserted at this site. By co-transfecting the packaging cells with multiple plasmids of the packaging component and the vector component, replication-defective lentiviral vector particles carrying the target gene can be harvested in the cell supernatant.

The lentiviral expression vector contains the genetic information required for packaging, transfection, and stable integration. The lentiviral packaging plasmid can provide all the auxiliary proteins required for transcription and packaging RNA into the recombinant pseudovirus vector. In order to produce high-titer viral particles, it is necessary to co-transfect the cells with the expression vector and the packaging plasmid at the same time. The virus is packaged in the cell, and the packaged pseudovirus particles are secreted into the culture medium outside the cell. After centrifugation to obtain the supernatant, it can be directly used for infection of host cells. After the target gene enters the host cell, it is reverse transcribed and integrated into the genome, thereby expressing the effector molecule at a high level.