Three types of Lentiviral Vectors - by GEG-tech.com

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Three types of Lentiviral Vectors - by GEG-tech.com

  1. 1. LentiVectors, EpiVectors, TransVectors Differences between the three cycles First let’s see how a classic Lentiviral vector works. © 2014 GEG-Tech, all rights reserved
  2. 2. LentiVectors After fusion with the cell membrane, the vector genome enters the cytoplasm. © 2014 GEG-Tech, all rights reserved
  3. 3. LentiVectors The RNA vector genome is then transcribed by the viral reverse-transcriptase (RT) into linear DNA. © 2014 GEG-Tech, all rights reserved
  4. 4. LentiVectors After translocation in the cell nucleus, the DNA is integrated into the cell chromatin by the HIV-1 integrase © 2014 GEG-Tech, all rights reserved
  5. 5. LentiVectors Using the cell machinery, your protein of interest is synthetized from the integrated vector genome © 2014 GEG-Tech, all rights reserved
  6. 6. LentiVectors With the integration of the vector genome, the protein is assured to have a high-level and long-term expression © 2014 GEG-Tech, all rights reserved
  7. 7. Now, let’s see how a non-integrating lentiviral vector works © 2014 GEG-Tech, all rights reserved
  8. 8. EpiVectors Beside integrating into the chromatin, the DNA of viral origin can also form circles… © 2014 GEG-Tech, all rights reserved
  9. 9. EpiVectors … which can be used as template for transcription. © 2014 GEG-Tech, all rights reserved
  10. 10. If we disable the viral integrase… © 2014 GEG-Tech, all rights reserved
  11. 11. EpiVectors The vector is no longer integrated but only circularized… © 2014 GEG-Tech, all rights reserved
  12. 12. EpiVectors … those circles are now the only template used for your protein synthesis. © 2014 GEG-Tech, all rights reserved
  13. 13. EpiVectors Due to the dilution effect of cell growth, these circles can provide short-term expression in dividing cells. © 2014 GEG-Tech, all rights reserved
  14. 14. EpiVectors However in non-dividing cells, you will have a long-term expression, yet lower than from integrated vectors. © 2014 GEG-Tech, all rights reserved
  15. 15. If you want transient expression of your transgene even in non-dividing cells, TransVectors are what you need. How does it work ? © 2014 GEG-Tech, all rights reserved
  16. 16. TransVectors In a classic lentiviral vector, the reversetranscriptase binds to a specific region of the vector genome called PBS © 2014 GEG-Tech, all rights reserved
  17. 17. TransVectors In our TransVectors, the PBS region is deleted from the vector genome. © 2014 GEG-Tech, all rights reserved
  18. 18. TransVectors The binding of RT and reverse-transcription are made impossible. © 2014 GEG-Tech, all rights reserved
  19. 19. TransVectors And without reverse-transcription, the whole cycle is impaired. © 2014 GEG-Tech, all rights reserved
  20. 20. TransVectors Your protein synthesis now rely on a very discreet process: translation directly from the primary RNA vector genome © 2014 GEG-Tech, all rights reserved
  21. 21. TransVectors Since RNAs have a short lifetime in the cytoplasm, the protein expression will be highly transient, even in non-dividing cells. © 2014 GEG-Tech, all rights reserved
  22. 22. LentiVectors, EpiVectors, TransVectors Differences between the three cycles The three lines of GEG-tech vectors provide you with the opportunity to delimitate your transgene expression in time. © 2014 GEG-Tech, all rights reserved
  23. 23. Viral genome final state LentiVectors EpiVectors TransVectors Transgene expression level Integrated DNA Transgene expression in: Dividing cells Non-dividing cells High Long term Long term Episomal DNA Medium Transient Long term RNA Low Transient Transient © 2014 GEG-Tech, all rights reserved

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