Synthesis, Spectroscopic Studies and Applications of
23rd IUPAC Conference on Physical Organic Chemistry (ICPOC23) 3rd – 8th July 2016 • Sydney • Australia
Synthesis, Spectroscopic Studies and Applications of Novel Cyanine-Styryl Dyes C. Schwechheimera,* and H.-A. Wagenknechta a
Institute of Organic Chemistry, Karlsruhe Institute of Technology, Karlsruhe, Germany *Correspondence to: [email protected]
% of Fluorescence F0
MicroRNAs (miRNA, miR) are single stranded non-coding oligonucleotides (19-24 nt) which play an important role in many biological processes, e. g. RNA interference (RNAi).1 Therefore miRNA-imaging offers valuable information to miRNA-involving cellular developments and diseases like cancer (oncomir). Low transfection of fluorescent probes, low miRNA expression and poor optical properties of fluorophores mainly limit the fluorescence brightness cells.2 Furthermore a high photostability enables long-term observations and reduces the phototoxicity.3-5 Cyanine-styryl dyes, like CyIQ and 1, are suitable tools for nucleic acid probes due to their high fluorescence intensity enhancement in presence of nucleic acids, an outstanding optical behavior and an improved photostability.6,7 100 80 60
1 CyIQ
40 20 0
0
60
120 180 240 300 360 420 480
Irradiation time t / min
Figure 1: Structure, quantum yields ΦF, half-life times t1/2 and photostabilities of the dyes 1 and CyIQ (grey structure). Photostabilities were determined by measuring the loss of fluorescence intensity F during irradiation with an 470nm-LED of the click-modified DNA single strands (2.5 µM ssDNA, 10 mM NaPi, 250 mM NaCl).
In this study, the combination of synthetically optimized fluorophores and covalently bound cyclooctatetraene (1) enabled a significant increase in photostability by more than 300%, as well as far improved optical properties. Hereby novel, powerful and photostable fluorophores were developed for nucleic acid detection and opened up a wide range of applications.3,4 This will now be further investigated in hybridization probes to detect the oncomir miR-155-5p, which is commonly overexpressed in tumors, e. g. breast cancer.1 In all probability the overexpression is detectable by these hybridization probes and also long-term studies within the zerbrafish cancer model (Danio rerio) are planned. References 1. 2. 3. 4. 5. 6. 7.
Faraoni, I.; Antonetti, F. R; Cardone, J.; Bonmassar, E. BBA – Molecular Basis of Disease 2009, 1792, 497-505. Ko, H. Y.; Lee, J.; Joo, J. Y.; Lee, Y. S.; Heo, H.; Ko, J. J.; Kim, S. Sci. Rep. 2014, 4, 4626. Zheng, Q.; Juette, M. F.; Jockusch, S.; Wasserman, M. R.; Zhou, Z.; Altman, R. B.; Blanchard, S. C. Chem. Soc. Rev. 2014, 43, 1044-1056. Ha, T.; Tinnefeld, P. Annu. Rev. Phys. Chem. 2012, 63, 595-617. Dixit, R.; Cyr, R. Plant J. 2003, 36, 280-290. Rubner, M. M.; Holzhauser, C.; Bohländer, P. R.; Wagenknecht, H.-A. Chem. Eur. J. 2012, 18, 1299-1302. Bohländer, P. R.; Wagenknecht, H.-A. Org. Biomol. Chem. 2013, 11, 7458-7462.
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Synthesis, Spectroscopic Studies and Applications of Novel Cyanine-Styryl Dyes C. Schwechheimera,* and H.-A. Wagenknechta a
Institute of Organic Chemistry, Karlsruhe Institute of Technology, Karlsruhe, Germany *Correspondence to: [email protected]
% of Fluorescence F0
MicroRNAs (miRNA, miR) are single stranded non-coding oligonucleotides (19-24 nt) which play an important role in many biological processes, e. g. RNA interference (RNAi).1 Therefore miRNA-imaging offers valuable information to miRNA-involving cellular developments and diseases like cancer (oncomir). Low transfection of fluorescent probes, low miRNA expression and poor optical properties of fluorophores mainly limit the fluorescence brightness cells.2 Furthermore a high photostability enables long-term observations and reduces the phototoxicity.3-5 Cyanine-styryl dyes, like CyIQ and 1, are suitable tools for nucleic acid probes due to their high fluorescence intensity enhancement in presence of nucleic acids, an outstanding optical behavior and an improved photostability.6,7 100 80 60
1 CyIQ
40 20 0
0
60
120 180 240 300 360 420 480
Irradiation time t / min
Figure 1: Structure, quantum yields ΦF, half-life times t1/2 and photostabilities of the dyes 1 and CyIQ (grey structure). Photostabilities were determined by measuring the loss of fluorescence intensity F during irradiation with an 470nm-LED of the click-modified DNA single strands (2.5 µM ssDNA, 10 mM NaPi, 250 mM NaCl).
In this study, the combination of synthetically optimized fluorophores and covalently bound cyclooctatetraene (1) enabled a significant increase in photostability by more than 300%, as well as far improved optical properties. Hereby novel, powerful and photostable fluorophores were developed for nucleic acid detection and opened up a wide range of applications.3,4 This will now be further investigated in hybridization probes to detect the oncomir miR-155-5p, which is commonly overexpressed in tumors, e. g. breast cancer.1 In all probability the overexpression is detectable by these hybridization probes and also long-term studies within the zerbrafish cancer model (Danio rerio) are planned. References 1. 2. 3. 4. 5. 6. 7.
Faraoni, I.; Antonetti, F. R; Cardone, J.; Bonmassar, E. BBA – Molecular Basis of Disease 2009, 1792, 497-505. Ko, H. Y.; Lee, J.; Joo, J. Y.; Lee, Y. S.; Heo, H.; Ko, J. J.; Kim, S. Sci. Rep. 2014, 4, 4626. Zheng, Q.; Juette, M. F.; Jockusch, S.; Wasserman, M. R.; Zhou, Z.; Altman, R. B.; Blanchard, S. C. Chem. Soc. Rev. 2014, 43, 1044-1056. Ha, T.; Tinnefeld, P. Annu. Rev. Phys. Chem. 2012, 63, 595-617. Dixit, R.; Cyr, R. Plant J. 2003, 36, 280-290. Rubner, M. M.; Holzhauser, C.; Bohländer, P. R.; Wagenknecht, H.-A. Chem. Eur. J. 2012, 18, 1299-1302. Bohländer, P. R.; Wagenknecht, H.-A. Org. Biomol. Chem. 2013, 11, 7458-7462.
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