Tsien again!

'Mike' Michael J. Moser moser at U.WASHINGTON.EDU
Mon Jul 20 00:06:19 EST 1998

I agree it sounds pretty cool.  FITC signal will be much brighter than
that of GFP.

But I wonder what the physiological consequences of "tetra-Cys" fusion
are and how the compare to those of GFP fusion.  Also, FDG staining of
cells is difficult and can lead to high backgrounds, hopefully this new
compound won't suffer from that problem.

It definitely sounds like a great alternative with potential to be a real
improvement, especially if the emmision and binding properties of the
fluorchrome can be varied to allow simultaneous user of multiple probes! 


Mike Moser                                            Tel: 206-543-6585
UW Department of Pathology                            FAX: 206-543-3967
Box 357705                                       moser at u.washington.edu
Seattle, WA  98195                 http://weber.u.washington.edu/~moser

On 17 Jul 1998, A. S. Goustin Lab wrote:

> With all this talk about power supplies and stainless steel, I am
> wondering if I am in the right group.  Isn't anyone as blown away in awe
> as I am with Roger Tsien's latest coup:
> Griffin BA, Adams SR, Tsien RY.  (1998).  Specific covalent labeling of
>      recombinant protein molecules inside living cells.
>      Science 281, 269-273.
> He shows that one can add a little bit of alpha-helix (WEAAAREACCRECCARA)
> to a protein and use this as a target for the fluorescent reporter
> FLASH-EDT2.  The idea is to engineer the target as a tag for your favorite
> protein, transfect the cell with a construct expressing this tagged
> protein, and then feed the cell:
>            4',5'-bis(1,3,2-dithioarsolan-2-yl)fluorescein
> which converts to FLASH-EDT2.  The dye homes to the tagged protein and now
> you can visualize your tagged protein IN A LIVING CELL!  Without gfp!
> The FLASH-peptide complex is ~50,000x more fluorescent than FLASH alone.
> It seems like you can make your own FLASH-EDT2 according to note 20 on p
> 271, or you can wait for some vendor to sell it commercially.
> I smell a great discovery...
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