title: Green-to-red photoconvertible Dronpa mutant for multimodal super-resolution fluorescence microscopy.
abstract: Advanced imaging techniques crucially depend on the labels used. In this work, we present the structure-guided design of a fluorescent protein that displays both reversibly photochromic and green-to-red photoconversion behavior. We first designed ffDronpa, a mutant of the photochromic fluorescent protein Dronpa that matures up to three times faster while retaining its interesting photochromic features. Using a combined evolutionary and structure-driven rational design strategy, we developed a green-to-red photoconvertible ffDronpa mutant, called pcDronpa, and explored different optimization strategies that resulted in its improved version, pcDronpa2. This fluorescent probe combines a high brightness with low photobleaching and photoblinking. We herein show that, despite its tetrameric nature, pcDronpa2 allows for multimodal subdiffraction imaging by sequentially imaging a given sample using both super-resolution fluctuation imaging and localization microscopy.
authors.affiliation: Department of Chemistry, KU Leuven , Celestijnenlaan 200F, bus 2404, 3001 Heverlee, Belgium.
authors.forename: Benjamien
authors.initials: B
authors.lastname: Moeyaert
authors.forename: Ngan
authors.initials: N
authors.lastname: Nguyen Bich
authors.forename: Elke
authors.initials: E
authors.lastname: De Zitter
authors.forename: Susana
authors.initials: S
authors.lastname: Rocha
authors.forename: Koen
authors.initials: K
authors.lastname: Clays
authors.forename: Hideaki
authors.initials: H
authors.lastname: Mizuno
authors.forename: Luc
authors.initials: L
authors.lastname: van Meervelt
authors.forename: Johan
authors.initials: J
authors.lastname: Hofkens
authors.forename: Peter
authors.initials: P
authors.lastname: Dedecker