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Engineering GPCR signaling pathways with RASSLs

Nature Methods volume 5, pages 673–678 (2008)Cite this article

Abstract

We are creating families of designer G protein–coupled receptors (GPCRs) to allow for precise spatiotemporal control of GPCR signaling in vivo. These engineered GPCRs, called receptors activated solely by synthetic ligands (RASSLs), are unresponsive to endogenous ligands but can be activated by nanomolar concentrations of pharmacologically inert, drug-like small molecules. Currently, RASSLs exist for the three major GPCR signaling pathways (Gs, Gi and Gq). We review these advances here to facilitate the use of these powerful and diverse tools.

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Figure 1: Creating RASSLs by targeted mutagenesis.
Figure 2: Ligand-dependent and -independent phenotypes are induced by tissue-specific expression of a hRO-i, a Gi RASSL.
Figure 3: Directed molecular evolution to create new RASSLs.

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Acknowledgements

We thank our funding agencies and the members of our labs who have contributed to the RASSL projects: US National Institutes of Health (HL60664-07 to B.R.C.; DK072071 to R.A.N.; and U19MH82441 to B.L.R), a National Alliance for Research on Schizophrenia and Depression Distinguished Investigator Award (to B.L.R.), the American Heart Association pre-doctoral fellowship program (0415005Y to W.C.C.), the Veterans Affairs Merit Review and Career Scientist Program (R.A.N.) and the Gladstone–California Institute of Regenerative Medicine, fellowship program (grant T2-00003 to E.C.H.). The Gladstone Institutes received support from the US National Center for Research Resources Grant RR18928-01. We thank J. Ng, T. Nguyen, D. Srivastava, R.W. Mahley, H. Zahed, B. Phillips, M. Spindler, G. Howard and S. Ordway for providing valuable technical assistance and discussions.

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Authors and Affiliations

  1. Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, 94158, California, USA

    Bruce R Conklin, Edward C Hsiao, W C Chang & Robert A Nissenson

  2. Departments of Medicine and Cellular and Molecular Pharmacology, University of California, San Francisco, 505 Parnassus Avenue, Room M994, San Francisco, 94122, California, USA

    Bruce R Conklin & Edward C Hsiao

  3. and Departments of Medicine and Physiology, Endocrine Research Unit, Veterans Affairs Medical Center, University of California, San Francisco, 4150 Clement Street, San Francisco, 94121, California, USA

    Edward C Hsiao & Robert A Nissenson

  4. Centre National de la Recherche Scientifique, UMR 5203, Institut de Génomique Fonctionnelle, 141 rue de la Cardonille and Institut National de la Santé et de la Recherche Médicale, U661, and Université Montpellier, Montpellier, 34094, France

    Sylvie Claeysen, Aline Dumuis & Joël Bockaert

  5. Department of Physiology, 600 16th St. University of California, San Francisco, San Francisco, 94143, California, USA

    Supriya Srinivasan

  6. Department of Neurosurgery, University of California, San Francisco, MCB 226, 1855 Folsom Street, San Francisco, 94103, California, USA

    John R Forsayeth

  7. Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville Pike, Bethesda, 20892, Maryland, USA

    Jean-Marc Guettier & Jürgen Wess

  8. Graduate Program in Pharmaceutical Sciences and Pharmacogenomics, University of California, San Francisco, 1700 Fourth Street, Byers Hall, Suite BH-216, San Francisco, 94143, California, USA

    W C Chang

  9. Department of Pharmacology, University of North Carolina, Chapel Hill, 1106 Mary Ellen Jones Building, CB 7365, 98 Manning Drive, Chapel Hill, 27599, North Carolina, USA

    Ying Pei, Ken D McCarthy & Bryan L Roth

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  1. Bruce R Conklin
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Correspondence to Bruce R Conklin or Bryan L Roth.

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Conklin, B., Hsiao, E., Claeysen, S. et al. Engineering GPCR signaling pathways with RASSLs. Nat Methods 5, 673–678 (2008). https://doi.org/10.1038/nmeth.1232

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