Lentiviral vector construction is a key technology in modern biomedical research. This article provides a detailed explanation of the definition and characteristics of lentiviral vector construction, describes its construction process, and hopes to give readers a deeper understanding of lentiviral vector construction while enhancing their biological science knowledge.
Lentiviral vector construction refers to the use of molecular biology techniques to remove non-essential genes from the HIV-1 viral genome, retaining only the cis-acting elements required for packaging, reverse transcription, and integration. At the same time, a target gene or RNAi sequence is introduced to build a viral vector system capable of efficiently infecting and integrating into the host cell genome. Lentiviral vector construction technology has become an important tool in the fields of gene therapy, gene editing, and transgenic animals.
Efficient infection: Lentiviral vectors can effectively infect various types of cells, including both dividing and non-dividing cells such as neurons and stem cells, thus improving gene transduction efficiency.
Persistent expression: The target gene integrates into the host cell genome and can be stably inherited through cell division, achieving long-term expression.
Low immunogenicity: Most of the genes from the HIV-1 virus have been deleted, reducing the vector's immunogenicity and minimizing immune responses after injection.
High biosafety: Constructed through a multi-plasmid system, which further reduces the risk of reverting to a wild-type virus, thus enhancing the safety of the vector.
Plasmid construction: Lentiviral vector construction includes the transfer plasmid containing the target gene or RNAi, as well as the packaging plasmid and envelope plasmid that provide the proteins needed for viral packaging.
Cell transfection: Co-transfect the aforementioned plasmids into packaging cells (such as 293T cells) to allow the viral genome to replicate within the cells and be packaged into viral particles.
Virus collection and purification: Collect the viral supernatant from the cell culture medium and purify the viral particles through methods like ultracentrifugation.
Titer determination: Measure the titer of the virus through infection experiments to ensure the infectivity of the viral vector.
