Lenti/Retro:What happens if I.. - (Mar/28/2007 )

Hi, I really get confused often here with retro/lenti, stable/transient stuffs. WHat happens if :
I transfect my dividing cells (say X) with pBABE puro or a lentiviral vector with my gene of interest without any helper constructs. So without production of viruses is it possible upon transfection, when the cells devide the gene of interest cloned between the LTRs will get integrated stably or it will just be a episome and get diluted later????? When is it necessary to produce viruses and transduce them??

I dont know reasons others use viruses, but I use it mainly to transfect neurons, which are generally considered not very easy to transfect. Some people may say its easier to transfect neurons with some reagents. but what efficiencies are we talking about and how about the toxicity. I agree some viruses are toxic but many are not.

Also viruses are the best way to transfect different cell types in vivo. Things like gene therapy use viruses mostly, they need long term expression.

Well some viruses are used just for the reason that they can infect neurons very efficiently and give extremely high expression in 24 hrs. One cannot use these viruses for therapy, they are toxic.

With lenti viruses, its easy to make them compared to others like AAV or adeno virus or HSV. But labs do make all these viruses regularly.

if you transfect cells lines, you could make stables and play with them but one requires lot of time to make them comapred to making a virus.

You could without making virus use the plasmid to make stables else the cells will lose the DNA. This would work. If its eposiomally present, some time down the road the cell will lose the DNA.

Thanks Scolix