On 23 May 2001, Guohong liu wrote:
> Has anyone tried to establish a stable cell line with GFP constructs? Is
> it toxic to cells?
Stable transfection of other proetins have been sucessful
in this lab. Roger Tsien has a number of transgenic animals
expressing all kinds of fluorescent proteins and I have
not heard anything about the animals dying from the
fluorescent proteins. Finally, we have done countless
transient transfections on tumor cells and it seems that
the tranfection protocols themselves are toxic (lipofectamine,
etc), rather than the GFP, EGFP, YFP, CFP, or the co-expressed
proteins (pIRIS2 vector from Clontech).
One proviso: Assuming the toxicity does not actually kill
the cells during the course of your experiment, your measure
and definition of toxicity may be an issue. Consider the case
of caspase 3. It is possible in some cells to activate caspase 3
without killing the cells during an experiment, but I think it
is well-accepted that once you have activated caspase 3, sooner
or later the cells will dye by an irreversible apoptosic pathway.
It is only a matter of time. I can't comment on necrosis.
The above seems to indicate that there is no general tendency
for cell death in cells expressing GFP since there exist
healthy trangenic animals expressing this or similar fluorescent
proteins. Was there not a recent Science paper in which a
trangenic primate was produced expressing GFP or some related
fluorescent protein?
Cheers,
Dominic
North 59 37' 30"
East 17 48' 10"
