title: Live imaging of endogenous protein dynamics in zebrafish using chromobodies.
abstract: Chromobodies are intracellular nanoprobes that combine the specificity of antibodies with the convenience of live fluorescence imaging in a flexible, DNA-encoded reagent. Here, we present the first application of this technique to an intact living vertebrate organism. We generated zebrafish lines expressing chromobodies that trace the major cytoskeletal component actin and the cell cycle marker PCNA with spatial and temporal specificity. Using these chromobodies, we captured full localization dynamics of the endogenous antigens in different cell types and at different stages of development. For the first time, the chromobody technology enables live imaging of endogenous subcellular structures in an animal, with the remarkable advantage of avoiding target protein overexpression or tagging. In combination with improved chromobody selection systems, we anticipate a rapid adaptation of this technique to new intracellular antigens and model organisms, allowing the faithful description of cellular and molecular processes in their dynamic state.
authors.affiliation: Max-Planck-Institut für Entwicklungsbiologie, Abteilung Genetik, Spemannstraße 35, Tübingen 72076, Germany paolo.panza@tuebingen.mpg.de.
authors.forename: Paolo
authors.initials: P
authors.lastname: Panza
authors.affiliation: Naturwissenschaftliches und Medizinisches Institut der Universität Tübingen, Markwiesenstraße 55, Reutlingen 72770, Germany Pharmazeutische Biotechnologie, Eberhard Karls Universität Tübingen, Auf der Morgenstelle 8, Tübingen 72076, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Maier
authors.affiliation: Naturwissenschaftliches und Medizinisches Institut der Universität Tübingen, Markwiesenstraße 55, Reutlingen 72770, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Schmees
authors.affiliation: Naturwissenschaftliches und Medizinisches Institut der Universität Tübingen, Markwiesenstraße 55, Reutlingen 72770, Germany Pharmazeutische Biotechnologie, Eberhard Karls Universität Tübingen, Auf der Morgenstelle 8, Tübingen 72076, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Rothbauer
authors.affiliation: Max-Planck-Institut für Entwicklungsbiologie, Abteilung Genetik, Spemannstraße 35, Tübingen 72076, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Söllner
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002453:Cell Cycle / Q000502:physiology; D003952:Diagnostic Imaging / Q000379:methods*; D061905:Single-Domain Antibodies; D015027:Zebrafish / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000378:metabolism*
chemical_list: D000199:Actins; D061905:Single-Domain Antibodies; D029961:Zebrafish Proteins
keywords: Actin; Live imaging; Nanobody; PCNA; Protein dynamics