title: Rational design of a monomeric and photostable far-red fluorescent protein for fluorescence imaging in vivo.
abstract: Fluorescent proteins (FPs) are powerful tools for cell and molecular biology. Here based on structural analysis, a blue-shifted mutant of a recently engineered monomeric infrared fluorescent protein (mIFP) has been rationally designed. This variant, named iBlueberry, bears a single mutation that shifts both excitation and emission spectra by approximately 40 nm. Furthermore, iBlueberry is four times more photostable than mIFP, rendering it more advantageous for imaging protein dynamics. By tagging iBlueberry to centrin, it has been demonstrated that the fusion protein labels the centrosome in the developing zebrafish embryo. Together with GFP-labeled nucleus and tdTomato-labeled plasma membrane, time-lapse imaging to visualize the dynamics of centrosomes in radial glia neural progenitors in the intact zebrafish brain has been demonstrated. It is further shown that iBlueberry can be used together with mIFP in two-color protein labeling in living cells and in two-color tumor labeling in mice.
authors.affiliation: Department of Pharmaceutical Chemistry, Cardiovascular Research Institute, University of California - San Francisco, San Francisco, California.
authors.forename: Dan
authors.initials: D
authors.lastname: Yu
authors.affiliation: Department of Bioengineering and Therapeutic Science, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Institute of Human Genetics, University of California, San Francisco, California.|College of Life Sciences and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Dong
authors.affiliation: Department of Pediatrics, Departments of Neurology and Neurological Surgery, University of California - San Francisco, San Francisco, California.
authors.forename: William Clay
authors.initials: WC
authors.lastname: Gustafson
authors.affiliation: Institut Químic De Sarrià, Universitat Ramon Llull, via Augusta 390, Barcelona, 08017, Spain.
authors.forename: Rubén
authors.initials: R
authors.lastname: Ruiz-González
authors.affiliation: Institut De Biologie Structurale (IBS), University of Grenoble Alpes, CNRS, CEA, Grenoble, F-38044, France.
authors.forename: Luca
authors.initials: L
authors.lastname: Signor
authors.affiliation: Institut De Biologie Structurale (IBS), University of Grenoble Alpes, CNRS, CEA, Grenoble, F-38044, France.
authors.forename: Fanny
authors.initials: F
authors.lastname: Marzocca
authors.affiliation: Institut De Biologie Structurale (IBS), University of Grenoble Alpes, CNRS, CEA, Grenoble, F-38044, France.
authors.forename: Franck
authors.initials: F
authors.lastname: Borel
authors.affiliation: Howard Hughes Medical Institute, Department of Physiology, University of California - San Francisco, San Francisco, California.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Klassen
authors.affiliation: Department of Pharmaceutical Chemistry, Cardiovascular Research Institute, University of California - San Francisco, San Francisco, California.
authors.forename: Kalpana
authors.initials: K
authors.lastname: Makhijani
authors.affiliation: Institut De Biologie Structurale (IBS), University of Grenoble Alpes, CNRS, CEA, Grenoble, F-38044, France.|European Synchrotron Radiation Facility, Grenoble, F-38043, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Royant
authors.affiliation: Howard Hughes Medical Institute, Department of Physiology, University of California - San Francisco, San Francisco, California.
authors.forename: Yuh-Nung
authors.initials: YN
authors.lastname: Jan
authors.affiliation: Department of Pediatrics, Departments of Neurology and Neurological Surgery, University of California - San Francisco, San Francisco, California.|Helen Diller Family Comprehensive Cancer Center, University of California - San Francisco, San Francisco, California.
authors.forename: William A
authors.initials: WA
authors.lastname: Weiss
authors.affiliation: Department of Bioengineering and Therapeutic Science, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Institute of Human Genetics, University of California, San Francisco, California.
authors.forename: Su
authors.initials: S
authors.lastname: Guo
authors.affiliation: Department of Pharmaceutical Chemistry, Cardiovascular Research Institute, University of California - San Francisco, San Francisco, California.
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Shu
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004330:Drosophila / Q000648:ultrastructure; D005260:Female; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009369:Neoplasms / Q000175:diagnosis; D061848:Optical Imaging / Q000379:methods*; D038761:Photobleaching; D011487:Protein Conformation; D055550:Protein Stability; D015027:Zebrafish / Q000196:embryology; D000097573:Red Fluorescent Protein
chemical_list: D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: bacterial phytochrome; fluorescence imaging; fluorescent proteins; rational design