Researchers

Chemistry / TSIEN, ROGER Y.

Position

Professor of Pharmacology at the University of California, San Diego, School of Medicine and Professor of Chemistry and Biochemistry at the University of California, San Diego.

Head of the Tsien Laboratory, University of California, San Diego, CA, USA.

Research interests

Roger Tsien's fascination with colors has revolutionized the fields of cell biology and neurobiology by allowing scientists to peer inside living cells and watch the behavior of molecules in real time. He is renowned for developing colorful dyes to track the movement of calcium within cells and has genetically modified molecules that make jellyfish and corals glow, creating fluorescent colors in a dazzling variety of hues. These multicolored fluorescent proteins have been used by scientists worldwide to track where and when certain genes are expressed in cells or in whole organisms. Now, Tsien is building on his fluorescent protein work to develop a novel way to image and possibly even deliver specially targeted drugs to cancer tumors.

Tsien has always been drawn to pretty colors. "Your science should ideally feed the deeper parts of your personality, to provide some intrinsic pleasure to tie you over the inevitable periods of discouragement," he says. Tsien grew up among a number of engineers in his extended family, and even from a young age he seemed destined for a career in science. Childhood asthma often kept Tsien indoors, where he spent hours conducting chemistry experiments in his basement laboratory and was first exposed to the chemistry of pretty colors. At 16, he won top prize in the nationwide Westinghouse Talent Search. He later attended Harvard College on a National Merit Scholarship, graduating at age 20 with a degree in chemistry and physics.

As a graduate student at the University of Cambridge, Tsien worked to develop a better dye to track inside cells the levels of calcium, which plays a critical role in numerous physiological processes, including the regulation of nerve impulses, muscle contraction, and fertilization. At that time, measuring intracellular calcium was a laborious process and typically involved injecting a calcium-binding protein through the cell membrane, a technique that often damaged the very cells being studied. Using techniques of chemistry, Tsien developed organic dyes that twist when they bind calcium, dramatically changing the dyes' fluorescence, and he found a way to masquerade the dyes so they could pass through the cell membrane without having to be injected.

In the early 1990s, Tsien borrowed from jellyfish a molecule that glows, green fluorescent protein, and reengineered it to emit colors ranging from blue to yellow. The fluorescent proteins can be tagged to certain genes or specific proteins of interest. Using a light microscope, scientists can easily determine by their glow when and where the genes are activated or the proteins are expressed. Over the years, Tsien has expanded the color palette of fluorescent proteins to include oranges, reds, and purples. He has also developed a way to monitor the interactions of two proteins, each tagged with different hues of fluorescent proteins. "As a whole, fluorescent proteins have had a huge impact on many areas of biological sciences because they gave [scientists] a direct link from genes and DNA to something you can see inside a cell or inside any organism," Tsien says, while also acknowledging that other scientists initially discovered and cloned fluorescent proteins.

Tsien recently has set his sights on the imaging and treatment of cancer. He and his colleagues have built U-shaped peptide molecules to carry a payload—an imaging molecule or chemotherapy drug. The peptides are substrates for certain proteases, protein-cleaving enzymes that are exuded from tumor cells but rarely appear on normal cells. When the protease cleaves the bottom of the U, the two arms of the U are separated, unleashing one arm to drag the payload portion of the peptide into a neighboring cancer cell. "I've always wanted to do something clinically relevant in my career, if possible, and cancer is the ultimate challenge," Tsien says.

Prizes and awards

2008 Nobel Prize in Chemistry

Gairdner Foundation International Award

Wolf Prize in Medicine

American Chemical Society Award for Creative Invention

Heineken Prize for Biochemistry and Biophysics, Royal Netherlands Academy of Sciences

Keio Medical Science Prize, Keio University, Japan

Publications

Reviews

Stephen R. Adams & Roger Y. Tsien 2006 Imaging the influx of cell-penetrating peptides into the cytosol of individual cells. Chapter 29 in: Handbook of Cell-Penetrating Peptides, ed by Ulo Langel (2nd edition). CRC 505-512.

Amy E. Palmer and Roger Y. Tsien 2006 Measuring calcium signaling using genetically targetable fluorescent indicators. Nature Protocols, Vol 1: 1057-1065.

Ben N. G. Giepmans, Stephen R. Adams, Mark H. Ellisman & Roger Y. Tsien 2006 The fluorescent toolbox for assessing protein location and function. Science. 312: 217-224.

Zacharias, D.A., and Tsien, R.Y. Molecular biology and mutation of green fluorescent protein. Chapter 5 in: Green Fluorescent Protein: Properties, Applications, and Protocols, ed. by M. Chalfie and S.R. Kain (2nd edition). John Wiley & Sons. This article is listed in Medline as Methods Biochem. Anal. 2006; 47: 83-120

Tsien, R.Y. 2005. Breeding molecules to spy on cells, in The Harvey Lectures: Series 99, 2003-2004. Harvey Society, 2005

Nathan C. Shaner, Paul A. Steinbach, & Roger Y. Tsien. 2005 . A guide to choosing fluorescent proteins, Nature Methods, Vol. 2: 905 – 909

Tsien, R.Y. 2003 Imagining imaging's future. Nature Reviews Molecular Cell Biology 4: SS16-SS21

Zhang, J., Campbell, R.E., Ting, A. and Tsien, R.Y. 2002. Creating new fluorescent probes for cell biology. Nature Review Molecular Biology 3: 906-918.

Miyawaki, A., and Tsien, R.Y. 2000. Monitoring protein conformations and interactions by fluorescence resonance energy transfer between mutants of green fluorescent protein. Methods in Enzymology 327: 472-500.

Wu, M.M., Llopis, J., Adams, S., McCaffery, J.M., Teter, K., Kulomaa, M.S., Machen, T.E.,Moore,H.-P.H., and Tsien, R.Y. 2000 Studying organelle physiology with fusion protein-targeted avidin and fluorescent biotin conjugates. Methods in Enzymology.327: 546-564.

Griffin, B.A., Adams, S.R., Jones, J., and Tsien, R.Y. 2000. Fluorescent labeling of recombinant proteins in living cells with FlAsH 2001. Methods in Enzymology. 327: 565-578.

Zacharias, D.A., Baird, G.S., and Tsien, R.Y. 2000. Recent advances in technology for measuring and manipulating cell signals. Current Opinion in Neurobiology 10: 416-421.

Tsien, R.Y. 1999. Monitoring Cell Calcium. In Calcium as a Cellular Regulator, ed. by E. Carafoli and C. Klee. New York: Oxford University Press, pp. 28-54.

Tsien, R.Y. 1998. The Green Fluorescent Protein. Annu. Rev. Biochemistry 67: 509-544.

Tsien. R.Y. and Pozzan T. 1989. Measurement of Cytosolic Free Ca2+ with Quin2. Method in Enzymology. 172: 230-244.

Papers

Stephen R. Adams & Roger Y. Tsien 2008 Preparation of the membrane-permeant biarsenicals FlAsH-EDT2 and ReAsH-EDT2 for fluorescent labeling of tetracysteine-tagged proteins. Nature Protocols 3: 1527-1534

Michael Z. Lin, Jeffrey S. Glenn, and Roger Y. Tsien 2008 A drug-controllable tag for visualizing newly synthesized proteins in cells and whole animals. Proc. Natl. Acad. Sci. 105: 7744–7749

Nathan C Shaner, Michael Z Lin, Michael R McKeown, Paul A Steinbach, Kristin L Hazelwood, Michael W Davidson & Roger Y Tsien 2008 Improving the photostability of bright monomeric orange and red fluorescent proteins. Nature Methods 5, 545-551

Samuel Andrew Hires, Yongling Zhu, and Roger Y. Tsien 2008 Optical measurement of synaptic glutamate spillover and reuptake by linker optimized glutamate-sensitive fluorescent reporters. Proc. Natl. Acad. Sci. 105: 4411–4416

Brent R. Martin, Thomas J. Deerinck, Mark H. Ellisman, Susan S.Taylor, and Roger Y.Tsien - Isoform-Specific PKA Dynamics Revealed by Dye-Triggered Aggregation and DAKAP1α-Mediated Localizationin Living Cells. Chemistry & Biology 14, 1031–1042

Oded Tour, Stephen R. Adams, Rex A. Kerr, Rene M. Meijer, Terrence J. Sejnowski, Richard W. Tsien, and Roger Y. Tsien - Calcium Green FlAsH as a genetically targeted small-molecule calcium indicator. Nature Chemical Biology. 3: 423-431

Hui-wang Ai, Nathan C. Shaner, Zihao Cheng, Roger Y. Tsien, and Robert E. Campbell 2007 - Exploration of new chromophore structures leads to the identification of improved blue fluorescent proteins. Biochemistry, 46, 5904-5910

Hauser, C.T., and Tsien, R.Y. 2007 A hexahistidine-Zn2+-dye label reveals STIM1
surface exposure. Proc. Natl. Acad. Sci. 104: 3693–3697

Wang, L., and Tsien, R.Y. 2006. Evolving proteins in mammalian cells using somatic hypermutation. Nature Protocols 1: 1346-1350.

Timothy A. Dunn, Chih-Tien Wang, Michael A. Colicos, Manuela Zaccolo, Lisa M. DiPilato, Jin Zhang, Roger Y. Tsien, and Marla B. Feller 2006 Imaging of cAMP levels and protein kinase A activity reveals that retinal waves drive oscillations in second-messenger cascades. J Neuroscience 26(49): 12807–12815

Guido M. Gaietta, Ben N. G. Giepmans, Thomas J. Deerinck, W. Bryan Smith, Lucy Ngan, Juan Llopis, Stephen R. Adams, Roger Y. Tsien, and Mark H. Ellisman 2006 Golgi twins in late mitosis revealed by genetically encoded tags for live cell imaging and correlated electron microscopy. Proc. Natl. Acad. Sci. 103: 17777–17782

Jeffrey J. Saucerman, JinZhang, Jody C. Martin, Lili X. Peng, Antine E. Stenbit, Roger Y. Tsien, and Andrew D. McCulloch 2006 Systems analysis of PKA-mediated phosphorylation gradients in live cardiac myocytes. Proc. Natl. Acad. Sci. 103: 12923–12928

Xiaokun Shu, Nathan C. Shaner, Corinne A.Yarbrough, Roger Y. Tsien, and S. James Remington 2006 Novel chromophores and buried charges control color in mFruits. Biochemistry 45: 9639-9647

Amy E. Palmer, Marta Giacomello, Tanja Kortemme, Andrew Hires, Varda Lev-Ram, David Baker, and Roger Y. Tsien. 2006 Ca2+ Indicators based on computationally redesigned calmodulin-peptide pairs. Chemistry & Biology 13: 521–530

Jeremy R. Babendure, Jennie L. Babendure, Jian-Hua Ding and Roger Y. Tsien. 2006 Control of mammalian translation by mRNA structure near caps. RNA,12: 851–861.
Supplemental Material

Brent R. Martin, Ben N.G. Giepmans, Stephen R. Adams & Roger Y. Tsien. 2005 Mammalian cell-based optimization of the biarsenical-binding tetracysteine motif for improved fluorescence and affinity. Nature Biotechnol. 10:1308-1314.

Jin Zhang, Christopher J. Hupfeld, Susan S. Taylor, Jerrold M. Olefsky & Roger Y. Tsien. 2005 Insulin disrupts Beta-adrenergic signaling to protein kinase A adipocytes. Nature, 437: 569-573.

Hoffmann, C., Gaietta, G., Bünemann, M., Adams, S.R., Oberdorff-Maass, S., Behr, B., Vilardaga, J., Tsien, R.Y., Ellisman, M.H., and Lohse, M.J.
2005 A FlAsH-based FRET approach to determine G protein-coupled receptor activation in living cells, Nature Methods 2: 171-176

Yingxiao Wang1, Elliot L. Botvinick, Yihua Zhao, Michael W. Berns, Shunichi Usami, Roger Y. Tsien & Shu Chien 2005 Visualizing the mechanical activation of Src, Nature 434: 1040-1045

JC Goldstein, C Munoz-Pinedo, J-E Ricci, SR Adams, A Kelekar, M Schuler, RY Tsien and DR Green 2005 Cytochrome c is released in a single step during apoptosis, Cell Death and Differentiation 12: 453–462

Tao Jiang, Emilia S. Olson, Quyen T. Nguyen, Melinda Roy, Patricia A. Jennings, and Roger Y. Tsien 2004 Tumor imaging by means of proteolytic activation of cell-penetrating peptides. Proc. Natl. Acad. Sci. 101: 17867-17872. Supplementary Material

Nathan C. Shaner, Robert E. Campbell, Paul Steinbach, Ben N. G. Giepmans, Amy E. Palmer and Roger Y. Tsien. 2004 Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma fluorescent protein. Nature Biotech. 22: 1567-1572

Amy E. Palmer, Can Jin, John C. Reed and Roger Y. Tsien 2004 Bcl-2-mediated alterations in endoplasmic reticulum Ca2+ analyzed with an improved genetically encoded fluorescent sensor Proc. Natl. Acad. Sci.101: 17404-17409.

Lei Wang, W. Coyt Jackson, Paul A. Steinbach, and Roger Y. Tsien 2004 Evolution of new nonantibody proteins via iterative somatic hypermutation Proc. Natl. Acad. Sci.101: 16745-16749.

Toshiaki Furuta, Hiroko Takeuchi, Masahiro Isozaki, Yasuhiro Takahashi, Makoto Kanehara, Masazumi Sugimoto, Takayoshi Watanabe, Kousei Noguchi, Timothy M. Dore, Takashi Kurahashi, Michiko Iwamura, and Roger Tsien. 2004 Bhc-cNMPs as either Water -Soluble or Membrane-Permeant Photoreleasable Cyclic Nucleotides for both One- and Two-Photon Excitation. Chem. Biochem. 5: 1119-1128.

Colette T. Dooley, Timothy M. Dore, George T. Hanson, W. Coyt Jackson, S. James Remington, and Roger Y. Tsien 2004 Imaging Dynamic Redox Changes in Mammalian Cells with Green Fluorescent Protein Indicators J. Biol. Chem., May 2004; 279: 22284 - 22293

George T. Hanson, Robert Aggeler, Devin Oglesbee, Mark Cannon, Roderick A. Capaldi, Roger Y. Tsien, and S. James Remington 2004 Investigating Mitochondrial Redox Potential with Redox-sensitive Green Fluorescent Protein Indicators J. Biol. Chem., Mar 2004; 279: 13044 -13053.

Ju, W., Morishita, W., Tsui, J., Gaietta, G., Deerink, T.J., Adams, S.R., Garner, C.G., Tsien, R.Y., Ellisman, M.H., and Malenka, R.C. 2004 Activity-dependent regulation of dendritic sysnthesis and trafficking of AMPA receptors. Nat. Neurosci. 7: 244-253

Ray, P., De, A., Min, J-J., Tsien, R.Y. and Gambhir, S.S. 2004 Imaging tri-fusion multimodality reporter gene expression in living subjects. Cancer Res. 64: 1323-1330.

Lev-Ram, V., Mehta, S.B., Kleinfeld, D., and Tsien, R.Y. 2003 Reversing cerebellar long-term depression. Proc. Natl. Acad. Sci. 100: 15989-15993

Hasegawa,S., Jackson, W.C., Tsien, R.Y., and Rao, J-H. 2003 Imaging tetrahymena ribozyme splicing activity in single live mammalian cells Proc. Natl. Acad. Sci. 100: 14892–14896.

Tour,O., Meijer,R.M., Zacharias,D.A., Adams.S.R. and Tsien, R.Y. 2003 Genetically targeted chromophore-assisted light inactivation Nat.Biotech 21: 1505-1508.

Babendure, J.R., Adams,S.R.and Tsien, R.Y. 2003 Aptamers Switch on Fluorescence of Triphenylmethane Dyes. J. Am. Chem. Soc. 125: 14716-14717.

Gao, W., Xing, B., Tsien, R.Y., and Rao, J-H. 2003. Novel fluorogenic substrates for imaging B-Lactamase Gene Expression. J.Am.Chem.Soc. 125: 11146-11147.

Violin, J.D., Zhang, J., Tsien, R.Y., and Newton, A.C. 2003. A genetically encoded fluorescent reporter reveals oscillatory phosphorylation by protein kinase C. J. Cell Biol. 161: 899-909.

Tsai, P.S., Friedman, B., Ifarraguerri, A.I., Thompson, B.D., Lev-Ram, V., Schaffer, C.B., Xiong, Q., Tsien, R.Y., Squier, J.A., and Kleinfeld, D. 2003. All-optical histology using ultrashort laser pulses. Neuron 39: 27-41

Yu D, Baird GS, Tsien RY, Davis RL. 2003 Detection of calcium transients in Drosophila mushroom body neurons with camgaroo reporters. J Neuroscience 23: 64-72.

Lev-Ram, V., Wong, S.T., Storm, D.R., and Tsien, R.Y. 2002 A new form of cerebellar long-term potentiation is postsynaptic and depends on nitric oxide but not cAMP. Proc. Natl. Acad. Sci. 99: 8389-8393.

Campbell, R.E., Tour, O., Palmer, A.E., Steinbach, P., Baird, G.S., Zacharias, D.A., and Tsien, R.Y. 2002. A monomeric red fluorescent protein. Proc. Natl. Acad. Sci. 99: 7877-7882.

Adams, S.R., Campbell, R.E., Gross, L.A., Martin, B.R., Walkup, G.K., Yao, Y., Llopis, J., and Tsien, R.Y. 2002. New biarsenical ligands and tetracysteine motifs for protein labeling in vitro and in vivo: synthesis and biological applications. J. Am. Chem. Soc., 124: 6063-6076.

Zacharias,D.A., Violin,J.D., Newton,A.C., Tsien R.Y. 2002 Partitioning of Lipid-Modified Monomeric GFPs into Membrane Microdomains of Live Cells Science 296: 913-916.

Gaietta, G., Deerinck, T.J., Adams, S.R., Bouwer, J., Tour,O., Laird, D.W., Sosinsky, G.E., Tsien, R.Y. and Ellisman, M.H. 2002 Multicolor and Electron Microscopic Imaging of Connexin Trafficking. Science 296: 503-507.

Zhang, J., Ma, Y., Taylor, S.S., and Tsien, R.Y. 2001 Genetically encoded reporters of protein kinase A activity reveal impact of substrate tethering Proc. Natl. Acad. Sci. 98: 14997-15002.

Ting, A.Y., Kain, K.H., Klemke, R.L. and Tsien, R.Y. 2001 Genetically encoded fluorescent reporters of protein tyrosine kinase activities in living cells Proc.Natl. Acad. Sci 98: 15003-15008.

Wu, M.M., Grabe, M., Adams, S., Tsien, R. Y., Moore, H-P. H., and Machen, T. E. 2001. Mechanisms of pH regulation in the regulated secretory pathway. J. Biol. Chem. 276: 33027-33035.

Burdette, S.C., Walkup, G.K., Spingler, B., Tsien, R.Y. and Lippard, S.J. 2001. Fluorescent sensors for Zn2+ based on a fluorescein platform: Synthesis, properties and intracellular distribution. J. Am. Chem. Soc. 123: 7831-7841.

Ando, H., Furuta, T., Tsien, R.Y., and Okamoto, H. 2001. Photo-mediated gene activation using caged RNA/DNA in zebrafish embryos. Nature Genetics 28: 317-325.

Chan, F, Siegel, R.M., Zacharias, D., Swofford, R., Holmes, K.L., Tsien, R.Y., Lenardo, M.J. (2001) Fluorescence Resonance Energy Transfer Analysis of Cell Surface Receptor Interactions and Signaling Using Spectral Variants of the Green Fluorescent Protein Cytometry. 44: 361-368

Griesbeck,O.; Baird,G.S.; Campbell,R.E.; Zacharias,D.A.; Tsien,R.Y. 2001 Reducing the environmental sensitivity of yellow fluorescent protein. mechanism and applications. J.Biol.Chem. 276: 29188-29194.

Honda, A., Adams, S.R., Sawyer, C.L., Lev-Ram, V., Tsien, R.Y., and Dostmann, W.R.G. 2001. Spatiotemporal dynamics of guanosine 3',5'-cyclic monophosphate revealed by a genetically encoded, fluorescent indicator. Proc. Natl. Acad. Sci., 98: 2437-2442.

Kapahi, P., Takahashi, T., Natoli, G., Adams, S.R., Chen, Y., Tsien, R.Y. and Karin, M. 2000. Inhibition of NF-κB activation by arsenite through reaction with a critical cysteine in the activation loop of IκB kinase. J. Biol. Chem. 275: 36062-36066.

Heikal, A.A., Hess, S.T., Baird, G.S., Tsien, R.Y., and Webb, W.W. 2000. Molecular Spectroscopy and Dynamics of Intrinsically Fluorescent Proteins: Coral Red (DsRed) and Yellow (Citrine). Proc. Natl. Acad. Sci., 97: 11996-12001

Gross, L.A., Baird, G.S., Hoffman, R.C., Baldridge, K.K., and Tsien, R.Y. 2000. The structure of the chromophore within DsRed, a red fluorescent protein from coral. Proc. Natl. Acad. Sci., 97: 11990-11995. Supplement

Baird, G.S., Zacharias, D.A., and Tsien, R.Y. 2000. Biochemistry, mutagenesis, and oligomerization of dsRed, a red fluorescent protein from coral. Proc. Natl. Acad. Sci., 97: 11984-11989.

Allen, G.J., Chu, S.P., Schumacher, K., Shimazaki, C.T., Vafeados, D., Kemper, A., Hawke, S.D., Tallman, G., Tsien, R.Y., Harper, J.F., Chory, J., and Schroeder, J.I. 2000. Alteration of stimulus-specific guard cell calcium oscillations and stomatal closing in Arabidopsis det3 mutant. Science 289: 2338-2342.

Siegel, R.M., Frederiksen, J.K., Zacharias,D.A., Chan, F.K.-M., Johnson, M., Lynch, D., Tsien, R.Y., and Lenardo, M.J. 2000. Fas preassociation required for apoptosis signaling and dominant inhibition by pathogenic mutations. Science 288: 2354-2357.

Kerr, R., Lev-Ram, V., Baird, G., Vincent, P., Tsien, R.Y., Schafer, W.R. 2000. Optical imaging of calcium transients in neurons and pharyngeal muscle of C. elegans. Neuron 26: 583-594.

Walkup, G.K., Burdette, S.C., Lippard, S.J., and Tsien, R.Y. 2000. A new cell-permeable fluorescent probe for Zn2+. J. Amer. Chem. Soc. 122: 5644-5645.

Matsuyama, S., Llopis, J., Deveraux, Q.L., Tsien, R.Y., and Reed, J.C. 2000. Changes in intramitochondrial and cytosolic pH: early events that modulate caspase activation during apoptosis. Nature Cell Biology 2: 318-325.

Llopis, J., Westin, S., Ricote, M., Wang, J., Cho, C.Y., Kurokawa, R., Rose, D.W., Rosenfeld, M.G., Tsien, R.Y., and Glass, C.K. 2000. Ligand-dependent interactions of coactivators SRC-1 and PBP with nuclear hormone receptors can be imaged in live cells and are required for transcription. Proc. Natl. Acad. Sci. USA 97: 4363-4368.

Wu, M.M., Llopis, J., Adams, S., McCaffery, J.M., Kulomaa, M.S., Machen, T.E., Moore, H.-P.H., and Tsien, R.Y. 2000. Organelle pH studies using targeted avidin and fluorescein-biotin. Chemistry & Biology 7: 197-209.

Zaccolo, M., De Giorgi, F., Cho, C.Y., Feng, L., Knapp, T., Negulescu, P.A., Taylor, S.S., Tsien, R.Y., and Pozzan, T. 2000. A genetically encoded, fluorescent indicator for cAMP in living cells. Nature Cell Biology 2: 25-29.

Thomas, D., Lipp, P., Tovey, S.C., Berridge, M.J., Li, W., Tsien, R.Y., and Bootman, M.D. 2000. Microscopic properties of elementary Ca2+ release sites in non‑excitable cells. Current Biology 10: 8-15.