I think I may have misunderstood something. If I transfect some premade stable packaging cell line with some expression vector will it stably produce lentiviral particles which will integrate whatever DNA is on the expression vector or will it only transiently express such particles and then revert back to producing only the default proteins of the packaging cell line. For example lets say I have the stable cell line which is expressing varius proteins needed for lentiviral particle assembly (ie the envelope, integrase, etc) I then transfect it with an expression vector for GFP. Now it should for at least a few days produce lentiviral particles which will integrate GFP into whatever cells are transduced with it. But will the packaging cell line continue producing such lentiviral particles (ie containing capable of integrating GFP) or will the cell line eventually stop expressing the information on the expression vector?
On Tuesday, December 11, 2012 12:14:48 AM UTC-5, Josiah Zayner wrote:
What are you trying to do?--Why not just make stable cell line?
On Monday, December 10, 2012 10:05:06 PM UTC-6, x11...@mail.com wrote:Alright this is a shot in the dark but if anyone can understand what I'm talking about and knows a company that sells custom stable lentiviral packaging cell lines (293t cells) which have already been transfected with some expression vector of a customers choosing please let me know. I know there are some that sell cell lines with lentiviral packaging vectors but can't find any that will transfect an expression vector in for me. The problem is that it's sorta easy to transfect cell lines will an expression vector and lentiviral packaging plasmids but this only results in transient lentiviral particle expression, that is to say the cells stop producing lentivirus particles after a few days. It is possible to make stable packaging cell lines it's just really hard (I can't do it on my own). I need a cell line that will produce lentivirus particles (which then can be used to integrate a gene of my choosing in other cells) for a long time (several months at least). So if anyone knows a company that sells or will make such a cell line please post a response.
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