Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it>

• Main Authors: Skourides Paris A, Andreou Maria, Charalambous Anna
• Format: Article
• Language: English
• Published: BMC 2009-12-01
• Series: Journal of Nanobiotechnology
• Online Access: http://www.jnanobiotechnology.com/content/7/1/9

<p>Abstract</p> <p>We describe an intein based method to site-specifically conjugate Quantum Dots (QDs) to target proteins <it>in vivo</it>. 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 (I_N). The C-terminus half (I_C) of the intein was conjugated to QDs <it>in vitro</it>. I_C-QD's and RNA encoding PH-I_Nwere microinjected into Xenopus embryos. <it>In vivo </it>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 <it>in vivo </it>methodology for the site-specific conjugation of QD's and other artificial structures to target proteins in different intracellular compartments and signaling complexes.</p>