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 |
| _version_ | 1798038371006676992 |
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| author | Skourides Paris A Andreou Maria Charalambous Anna |
| author_facet | Skourides Paris A Andreou Maria Charalambous Anna |
| author_sort | Skourides Paris A |
| collection | DOAJ |
| description | <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> |
| first_indexed | 2024-04-11T21:39:17Z |
| format | Article |
| id | doaj.art-4b5065dbf5204fe48f7e52de2041aef0 |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-11T21:39:17Z |
| publishDate | 2009-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-4b5065dbf5204fe48f7e52de2041aef02022-12-22T04:01:39ZengBMCJournal of Nanobiotechnology1477-31552009-12-0171910.1186/1477-3155-7-9Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it>Skourides Paris AAndreou MariaCharalambous Anna<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>http://www.jnanobiotechnology.com/content/7/1/9 |
| spellingShingle | Skourides Paris A Andreou Maria Charalambous Anna Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it> Journal of Nanobiotechnology |
| title | Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it> |
| title_full | Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it> |
| title_fullStr | Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it> |
| title_full_unstemmed | Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it> |
| title_short | Intein-mediated site-specific conjugation of Quantum Dots to proteins <it>in vivo</it> |
| title_sort | intein mediated site specific conjugation of quantum dots to proteins it in vivo it |
| url | http://www.jnanobiotechnology.com/content/7/1/9 |
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