Hi, I am a master's degree student, and I am doing gene silencing with miRNA. The way I have done with transfection was the lentiviral system transcribed my interested miRNA and got very good regulating efficiency. But if I want to observe that a higher level of miRNA (higher than the lentiviral system expressed ) could possibly result, then is it reasonable to directly transfect my interested siRNA mimic to the cell that has been transfected with the lentiviral vector
Actually, I have done this idea and got an even better result. But I was challenged by my advisor. She asked me where was your idea from and, were there other papers that have done this before?
I have done searching related papers on the Net, but I didn't find any similar method to prove my idea.
Dear Che-Wei Liu
Go ahead with your ideas use some common liposomal mediated transfection modify for your work and develop new method.
I think it depends on how much the cell can uptake. Now essentially the LV vector should be contained in the cells after transfection for awhile whereas the siRNA needs to be retreated every 3 or so days. I would try it, but do not do the transfection at the same time, or else you risk the cells taking up more lentivirus particles than siRNA or vice-versa. I have used an adenovirus vector and transfected with a luciferase reporter 48hours later and have also co-transfected 2 siRNA. Also be mindful of how to prove your cells are taking up both the LV particles and the siRNA.
Both (lentivirus and liposome) are vectors to transfer nucleic acid into the cells; why there is a need to co-transfect both? If both transfections are performed, how the transfection efficiency will be calculated?
If you want to perform such experiment, compare individual and co-transfection efficiency and silencing/inhibition effect.
Note: miRNA and siRNA mechanisms are completely different from each other; so interpretation of observation should be made with caution.
- Badges
- Science method