@article {65769,
	title = {Improved Chemical-Genetic Fluorescent Markers for Live Cell Microscopy},
	journal = {Biochemistry},
	volume = {57},
	year = {2018},
	month = {11/2018},
	pages = {5648 {\textendash} 5653},
	abstract = {<p>Inducible chemical-genetic fluorescent markers are promising tools for live cell imaging requiring high spatiotemporal resolution and low background fluorescence. The fluorescence-activating and absorption shifting tag (FAST) was recently developed to form fluorescent molecular complexes with a family of small, synthetic fluorogenic chromophores (so-called fluorogens). Here, we use rational design to modify the binding pocket of the protein and screen for improved fluorescence performances with four different fluorogens. The introduction of a single mutation results in improvements in both quantum yield and dissociation constant with nearly all fluorogens tested. Our improved FAST (iFAST) allowed the generation of a tandem iFAST (td-iFAST) that forms green and red fluorescent reporters 1.6-fold and 2-fold brighter than EGFP and mCherry, respectively, while having a comparable size.</p>
},
	doi = {10.1021/acs.biochem.8b00649},
	url = {https://pubs-acs-org.accesdistant.sorbonne-universite.fr/doi/pdf/10.1021/acs.biochem.8b00649},
	author = {Tebo, Alison G and Pimenta, Frederico M and Zhang, Yu and Gautier, Arnaud}
}
