How does lentiviral transduction work?
Principle: Lentiviruses—a subclass of retroviruses—have the ability to permanently integrate into the genome of the host cell. After the virus has entered the cell, the viral RNA is transcribed by the reverse transcriptase to produce double-stranded DNA that enters the nucleus.
How do you increase lentiviral transduction efficiency?
10 best practices for a good transduction efficiency
- 1- Know your cells.
- 2- Check the quality of your lentiviral vector batch.
- 3- Optimize the transgene expression cassette.
- 4- Make sure your titer is in TU/ml.
- 5- Follow storage and thawing requirements.
- 6- Monitor cells sensitivity towards transduction enhancers.
How much polybrene do I add?
Add 0.5 mL of a single viral dilution to each well (each well gets one dilution, so a 6-well plate will hold 5 dilutions plus one ‘no virus’ control well)….Procedure.
| Dilution | Volume of Lentivirus (μL) | Volume of DMEM complete + 10 µg/mL polybrene (µL) |
|---|---|---|
| 1:5 | 300 | 200 |
| 1:10 | 150 | 350 |
| 1:50 | 30 | 470 |
| 1:100 | 15 | 485 |
What is adenovirus transduction?
Adenoviral gene transfer is one of the most reliable methods for introducing genes into mammalian cells. Because infection by adenovirus is not cell-cycle dependent, you can deliver your gene to primary as well as transformed cell lines.
What is the difference between transfection and transduction?
Transfection is the process of introducing nucleic acids into cells by non-viral methods. Transduction is the process whereby foreign DNA is introduced into another cell via a viral vector.
How long can lentivirus be stored at 4c?
Storage of Lentivirus Virus can be stored at 4°C for a short time (less than a week) before using after reception. Since Lentiviruses are sensitive to freeze-thawing and the titer drops with repeated freeze-thawing, aliquot viral stock should be stored at – 80°C freezer immediately upon arrival for long-term usage.