Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it>
<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 suc...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2009-12-01
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| Series: | Journal of Nanobiotechnology |
| Online Access: | http://www.jnanobiotechnology.com/content/7/1/9 |
| Summary: | <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<sub>N</sub>). The C-terminus half (I<sub>C</sub>) of the intein was conjugated to QDs <it>in vitro</it>. I<sub>C</sub>-QD's and RNA encoding PH-I<sub>N </sub>were 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> |
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| ISSN: | 1477-3155 |