EF1α-Citrine-Puro Lentivirus

Lentivirus (LV) belongs to the Retroviridae family and is a spherical enveloped virus with a diameter of about 80-120nm. Its genome is a single-stranded positive-strand RNA containing two copies, contained in a core composed of enzymatic proteins nucleocapsid (NC), capsid (CA), reverse transcriptase (RT), integrase (IN) and protease (PR), and the core is surrounded by a protein layer of matrix protein (MA). The envelope protein (ENV) is embedded in the lipid envelope of the virus particle, which binds to the target cell receptor and mediates the entry of the virus particle. At both ends of the genome are long terminal repeats (LTRs), which enable the lentiviral genome to integrate into the host genome where transcription is more active, and it contains strong promoters and enhancers. So if we artificially replace the sequence between the LTRs with our target gene sequence, during the viral infection process, after the lentiviral genome enters the cell, it is reverse transcribed into DNA by the reverse transcriptase it carries in the cytoplasm to form a DNA pre-integration complex. After entering the cell nucleus, the DNA is integrated into the cell genome. The integrated DNA transcribes mRNA and returns to the cytoplasm to express the target protein or produce RNAi interference. It can be passed to daughter cells as the host cell divides and can also be stably expressed in the cell. In some cell-related experiments, some cells are difficult to transfect using conventional methods. Usually, the target gene is randomly integrated into the host genome by lentiviral infection, thereby increasing the transfection efficiency, and can be stably expressed in the cell line for several generations, and stable cell lines can be screened. At present, lentivirus has been widely used in gene function research, RNA interference, small molecule RNA research and live animal experiments, and has become one of the powerful tools for gene therapy.