Hi
I m looking for destabilized GFP (deGFP) expressing cell line or vectors (viral vectors to make stable expressing cell line).
I need this cell line to study the release kinetics of scaffold.
i would really appreciate the help, thanks
Dear Karthikeyan,
I have used a GFP:PEST protein (a destalibilised version of GFP with fast turnover) in past. The advantage of this destabilized protein is that if you put your GFP under the influence of your desired promoter, when this promoter is not active, the mRNA level of this fusion product goes down and so your destabilized protein. The kinetic of degradation is much faster than a conventional GFP so you can follow this more directly. This sort of protein are also essential when you want to measure some effector/treatments (i.e. reporter cell line ±drugs) and your system sufferes of counterselection (i.e. you activate/reperss something that the cells need.). You can read/measure your output before the cells start to die. I have the plasmid if you want. You may generate stable cell lines from it or further modify it. You may also have to ask the person that gave it to me for perimission or MTA.
Dear Andrea
Thanks for the prompt response.
I m checking siRNA (siGFP) released from scaffold on different days are functional. For this assay, i need a Destabilized GFP. I have done the assay with normal eGFP, there is no change in the expression (only change in the intensity, but no change in % of GFP positive cells in FACS).
It will be really helpful to my study if you could share the plasmids. I will write to the concern person for the MTA/Permission. Will you please give me the contact details of the person.
Thanks again, I really appreciate you help.
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