Kienzler and coworkers report a new FRET method using three chromophores instead of the conventional two. By adding three chromophores, multiplex systems could be probed as well as looking at longer distances compared to traditional FRET. This proof-of-principle paper used solid phase synthesis to introduce new building blocks with the chromophores attached.
I think it will be interesting to see where this technique goes. Could unnatural amino acids/nucleic acids be used to incorporate chromophores into cell?
The only thing that I am concerned with is that energy is lost during each transfer. Should you be concerned with whether you can get a signal from the third FRET pair?
ReplyDeleteWhat about the background? Doesn't the addition of a chromophore risk to increase background and non-specificity?
ReplyDeleteThe Schultz group just published a paper where they report incorporation of a fluorescent coumarin into cells via unnatural amino acid.
ReplyDeleteHere is the link if you want to check it out:
http://www.sciencedirect.com/science/article/pii/S0960894X11011528
Actually, the already reported the incorporation of that amino acid in 2006: http://pubs.acs.org/doi/abs/10.1021/ja062666k
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