title: Comparative study reveals better far-red fluorescent protein for whole body imaging.
abstract: Genetically encoded far-red and near-infrared fluorescent proteins enable efficient imaging in studies of tumorigenesis, embryogenesis, and inflammation in model animals. Here we report comparative testing of available GFP-like far-red fluorescent proteins along with a modified protein, named Katushka2S, and near-infrared bacterial phytochrome-based markers. We compare fluorescence signal and signal-to-noise ratio at various excitation wavelength and emission filter combinations using transiently transfected cell implants in mice, providing a basis for rational choice of optimal marker(s) for in vivo imaging studies. We demonstrate that the signals of various far-red fluorescent proteins can be spectrally unmixed based on different signal-to-noise ratios in different channels, providing the straightforward possibility of multiplexed imaging with standard equipment. Katushka2S produced the brightest and fastest maturing fluorescence in all experimental setups. At the same time, signal-to-noise ratios for Katushka2S and near-infrared bacterial phytochrome, iRFP720 were comparable in their optimal channels. Distinct spectral and genetic characteristics suggest this pair of a far-red and a near-infrared fluorescent protein as an optimal combination for dual color, whole body imaging studies in model animals.
authors.affiliation: Department of Radiology, University of Michigan Medical School, Ann Arbor, MI 48109-2200, USA.
authors.forename: K E
authors.initials: KE
authors.lastname: Luker
authors.affiliation: Tallinn University of Technology, Department of Gene Technology. 15 Akadeemia St, Tallinn 12618, Estonia.
authors.forename: P
authors.initials: P
authors.lastname: Pata
authors.affiliation: 1] Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Miklukho-Maklaya 16/10, 117997, Moscow, Russia [2] Evrogen JSC, Miklukho-Maklaya 16/10, 117997, Moscow, Russia.
authors.forename: I I
authors.initials: II
authors.lastname: Shemiakina
authors.affiliation: Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Miklukho-Maklaya 16/10, 117997, Moscow, Russia.
authors.forename: A
authors.initials: A
authors.lastname: Pereverzeva
authors.affiliation: Department of Radiology, University of Michigan Medical School, Ann Arbor, MI 48109-2200, USA.
authors.forename: A C
authors.initials: AC
authors.lastname: Stacer
authors.affiliation: 1] Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Miklukho-Maklaya 16/10, 117997, Moscow, Russia [2] Evrogen JSC, Miklukho-Maklaya 16/10, 117997, Moscow, Russia.
authors.forename: D S
authors.initials: DS
authors.lastname: Shcherbo
authors.affiliation: Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Miklukho-Maklaya 16/10, 117997, Moscow, Russia.
authors.forename: V Z
authors.initials: VZ
authors.lastname: Pletnev
authors.affiliation: Tallinn University of Technology, Department of Gene Technology. 15 Akadeemia St, Tallinn 12618, Estonia.
authors.forename: M
authors.initials: M
authors.lastname: Skolnaja
authors.affiliation: 1] Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Miklukho-Maklaya 16/10, 117997, Moscow, Russia [2] Nizhny Novgorod State Medical Academy, Nizhny Novgorod, Russia.
authors.forename: K A
authors.initials: KA
authors.lastname: Lukyanov
authors.affiliation: Department of Radiology, University of Michigan Medical School, Ann Arbor, MI 48109-2200, USA.
authors.forename: G D
authors.initials: GD
authors.lastname: Luker
authors.affiliation: Tallinn University of Technology, Department of Gene Technology. 15 Akadeemia St, Tallinn 12618, Estonia.
authors.forename: I
authors.initials: I
authors.lastname: Pata
authors.affiliation: 1] Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Miklukho-Maklaya 16/10, 117997, Moscow, Russia [2] CEITEC MU, Masaryk University, Brno, Czech republic [3] Pirogov Russian National Research Medical University, 117997 Moscow, Russia.
authors.forename: D M
authors.initials: DM
authors.lastname: Chudakov
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D057809:HEK293 Cells; D064593:Heterografts; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D023421:Models, Animal; D057054:Molecular Imaging / Q000379:methods; D022821:RNA Splice Sites; D059629:Signal-To-Noise Ratio; D051598:Whole Body Imaging / Q000379:methods; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D022821:RNA Splice Sites; D000097573:Red Fluorescent Protein