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

doi:10.1371/journal.pone.0075835

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

Biochemistry & Molecular Biology

Research output: Contribution to journal › Article › Research › peer-review

19 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 language	English
Article number	e75835
Number of pages	9
Journal	PLoS ONE
Volume	8
Issue number	9
DOIs	https://doi.org/10.1371/journal.pone.0075835
Publication status	Published - 2013

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10.1371/journal.pone.0075835Licence: CC BY

2321588-oaFinal published version, 1.81 MBLicence: CC BY

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title = "Phanta: A non-fluorescent photochromic acceptor for pcFRET",

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.",

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AU - Don Paul, Craig Bradley

AU - Kiss, Csaba

AU - Traore, Daouda A K

AU - Gong, Lan

AU - Wilce, Matthew Charles James

AU - Devenish, Rodney James

AU - Bradbury, Andrew

AU - Prescott, Mark

PY - 2013

Y1 - 2013

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AB - 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.

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Phanta: A non-fluorescent photochromic acceptor for pcFRET

Abstract

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