Q. How recombinant proteins are produced in mammalian cells?
The general process of producing a recombinant protein in a mammalian expression system starts with transfection of a recombinant gene integrated in a vector into the chosen cell. The gene carries structural elements that regulates transcription.
Q. What is lentivirus production?
Lentiviruses, such as the Human Immunodeficiency Virus, are capable to infecting both dividing and non-dividing cells. Lentiviral vector production systems are based on a ‘split’ system, where the natural viral genome has been split into individual helper plasmid constructs.
Q. Why are recombinant proteins produced in mammalian cells?
Cultivated mammalian cells have become the dominant system for the production of recombinant proteins for clinical applications because of their capacity for proper protein folding, assembly and post-translational modification.
Q. Where do lentiviruses integrate?
Where does lentivirus integrate? Genome-wide studies of viral integration have shown that lentiviruses most often integrate into actively transcribed genes, and that this preference is conserved across target species.
Q. How is lentiviral transduction used in mammalian cells?
Lentiviral transduction is an efficient method for the delivery of transgenes to mammalian cells and unifies the ease of use and speed of transient transfection with the robust expression of stable cell lines. In this protocol, we describe the design and step-by-step application of a lentiviral plasmid suite, termed pHR-CMV-TetO 2,…
Q. What are the survival rates of lentiviral vectors?
The TYF-CMV-eGFP-transduced cells began with a higher level of GFP expression than other vectors did. The percentage of GFP positive cells for any of the five lentiviral vectors sustained over time. Moreover, the survival rates of all transfected cells exceeded 80% at both 5 and 9 weeks post transduction.
Q. How are lentiviral particles produced in a cell line?
The procedure to transduce expression cells is straightforward; a lentivirus producer cell line (HEK293 Lenti-X) is transiently co-transfected with transfer (encoding the transgene of interest), envelope and packaging plasmids to generate lentiviral particles.
Q. Can a lentivirus be used for gene delivery?
The use of lentiviruses, belonging to the retrovirus genus, presents such a possibility. A recombinant lentivirus, containing genetic elements from HIV, is widely used for gene delivery in medical applications 5.
