title: Understanding the photophysics of the spinach-DFHBI RNA aptamer-fluorogen complex to improve live-cell RNA imaging.
abstract: The use of aptamer-fluorogen complexes is an emerging strategy for RNA imaging. Despite its promise for cellular imaging and sensing, the low fluorescence intensity of the Spinach-DFHBI RNA aptamer-fluorogen complex hampers its utility in quantitative live-cell and high-resolution imaging applications. Here we report that illumination of the Spinach-fluorogen complex induces photoconversion and subsequently fluorogen dissociation, leading to fast fluorescence decay and fluorogen-concentration-dependent recovery. The fluorescence lifetime of Spinach-DFHBI is 4.0 ± 0.1 ns irrespective of the extent of photoconversion. We detail a low-repetition-rate illumination scheme that enables us to maximize the potential of the Spinach-DFHBI RNA imaging tag in living cells.
authors.affiliation: Howard Hughes Medical Institute , Urbana, Illinois 61801, United States.
authors.forename: Kyu Young
authors.initials: KY
authors.lastname: Han
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Leslie
authors.forename: Jingyi
authors.initials: J
authors.lastname: Fei
authors.forename: Jichuan
authors.initials: J
authors.lastname: Zhang
authors.forename: Taekjip
authors.initials: T
authors.lastname: Ha
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000737:chemistry; D007700:Kinetics; D055668:Photochemical Processes; D018724:Spinacia oleracea / Q000737:chemistry*
chemical_list: D052157:Aptamers, Nucleotide; D049452:Green Fluorescent Proteins