Phanta: A non-fluorescent photochromic acceptor for pcFRET

Craig Bradley Don Paul, Csaba Kiss, Daouda A K Traore, Lan Gong, Matthew Charles James Wilce, Rodney James Devenish, Andrew Bradbury, Mark Prescott

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13 Citations (Scopus)

Abstract

We have developed an orange non-fluorescent photochromic protein (quantum yield, 0.003) we call Phanta that is useful as an acceptor in pcFRET applications. Phanta can be repeatedly inter-converted between the two absorbing states by alternate exposure to cyan and violet light. The absorption spectra of Phanta in one absorbing state shows excellent overlap with the emission spectra of a number of donor green fluorescent proteins including the commonly used EGFP. We show that the Phanta-EGFP FRET pair is suitable for monitoring the activation of caspase 3 in live cells using readily available instrumentation and a simple protocol that requires the acquisition of two donor emission images corresponding to Phanta in each of its photoswitched states. This the first report of a genetically encoded non-fluorescent acceptor for pcFRET.
Original languageEnglish
Article numbere75835
Number of pages9
JournalPLoS ONE
Volume8
Issue number9
DOIs
Publication statusPublished - 2013

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