Hi I am new to stable transfection and I am confused about the concept of "all-in-one" lentiviral vector. My goal is to generate a stable HEK293T cell line producing my gene of interest under the control of doxycycline.
In some protocols using "all-in-one" pCW57 lentiviral vector, packaging plasmids were co-transfected with pCW57. But in another protocol I found, the author did not use any packaging plasmids because pCW57 is already an "all-in-one" vector.
I would appreciate any opinions! Thanks all.
Hi Shan,
If I understand well your question, the mention "all in one" is about the plasmid which encodes the LV genome and not about the method of production of the lenti. That's mean that all the elements of regulation are in the plasmid of genome, but not that you can produce lenti with just this plasmid. To produce LVs, you generally do tri-transfection (for a question of biosafety): transcomplementation plasmid+envelope plasmid+genome plasmid
Hope this will help you
Regards
Nicolas
- Badges
- Science method