title: Intein-mediated site-specific conjugation of Quantum Dots to proteins in vivo.
abstract: We describe an intein based method to site-specifically conjugate Quantum Dots (QDs) to target proteins in vivo. This approach allows the covalent conjugation of any nanostructure and/or nanodevice to any protein and thus the targeting of such material to any intracellular compartment or signalling complex within the cells of the developing embryo. We genetically fused a pleckstrin-homology (PH) domain with the N-terminus half of a split intein (IN). The C-terminus half (IC) of the intein was conjugated to QDs in vitro. IC-QD's and RNA encoding PH-IN were microinjected into Xenopus embryos. In vivo intein-splicing resulted in fully functional QD-PH conjugates that could be monitored in real time within live embryos. Use of Near Infra Red (NIR)-emitting QDs allowed monitoring of QD-conjugates within the embryo at depths where EGFP is undetectable demonstrating the advantages of QD's for this type of experiment. In conclusion, we have developed a novel in vivo methodology for the site-specific conjugation of QD's and other artificial structures to target proteins in different intracellular compartments and signaling complexes.
authors.affiliation: Department of Biological Sciences, University of Cyprus, P,O, Box 20537, 1678 Nicosia, Cyprus. skourip@ucy.ac.cy.
authors.forename: Anna
authors.initials: A
authors.lastname: Charalambous
authors.forename: Maria
authors.initials: M
authors.lastname: Andreou
authors.forename: Paris A
authors.initials: PA
authors.lastname: Skourides