Phanta: a non-fluorescent photochromic acceptor for pcFRET.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 24098733.
- Also identified by DOI 10.1371/journal.pone.0075835 and PMC identifier 3786930.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Biosensing Techniques
- Fluorescence Resonance Energy Transfer
- Green Fluorescent Proteins
- Photochemistry
- Recombinant Proteins