Ruthenium-caged antisense morpholinos for regulating gene expression in zebrafish embryos† †Electronic supplementary information (ESI) available: Synthetic protocols and characterization data for RuBEP, Ru–DNA and Ru-MO, purification and injection protocols, and oligo sequences. CCDC 1041125. For ESI and crystallographic data in CIF or other electronic format see DOI: 10.1039/c4sc03990d Click here for additional data file. Click here for additional data file
| Autor: | Griepenburg, Julianne C., Rapp, Teresa L., Carroll, Patrick J., Eberwine, James, Dmochowski, Ivan J. |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: | |
| Zdroj: | Chemical Science |
| ISSN: | 2041-6539 2041-6520 |
| Popis: | Ruthenium photolinkers provide a versatile method of using visible light to control structure and function of biopolymers. Photochemical approaches afford high spatiotemporal control over molecular structure and function, for broad applications in materials and biological science. Here, we present the first example of a visible light responsive ruthenium-based photolinker, Ru(bipyridine)2(3-ethynylpyridine)2 (RuBEP), which was reacted stoichiometrically with a 25mer DNA or morpholino (MO) oligonucleotide functionalized with 3′ and 5′ terminal azides, via Cu(i)-mediated [3+2] Huisgen cycloaddition reactions. RuBEP-caged circular morpholinos (Ru-MOs) targeting two early developmental zebrafish genes, chordin and notail, were synthesized and tested in vivo. One-cell-stage zebrafish embryos microinjected with Ru-MO and incubated in the dark for 24 h developed normally, consistent with caging, whereas irradiation at 450 nm dissociated one 3-ethynylpyridine ligand (Φ = 0.33) and uncaged the MO to achieve gene knockdown. As demonstrated, Ru photolinkers provide a versatile method for controlling structure and function of biopolymers. |
| Databáze: | OpenAIRE |
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