Double-labeled donor probe can enhance the signal of fluorescence resonance energy transfer (FRET) in detection of nucleic acid hybridization

@article{citeulike:2524525, abstract = {A set of fluorescently-labeled DNA probes that hybridize with the target RNA and produce fluorescence resonance energy transfer (FRET) signals can be utilized for the detection of specific RNA. We have developed probe sets to detect and discriminate single-strand RNA molecules of plant viral genome, and sought a method to improve the FRET signals to handle in vivo applications. Consequently, we found that a double-labeled donor probe labeled with Bodipy dye yielded a remarkable increase in fluorescence intensity compared to a single-labeled donor probe used in an ordinary FRET. This double-labeled donor system can be easily applied to improve various FRET probes since the dependence upon sequence and label position in enhancement is not as strict. Furthermore this method could be applied to other nucleic acid substances, such as oligo RNA and phosphorothioate oligonucleotides (S-oligos) to enhance FRET signal. Although the double-labeled donor probes labeled with a variety of fluorophores had unexpected properties (strange UV-visible absorption spectra, decrease of intensity and decay of donor fluorescence) compared with single-labeled ones, they had no relation to FRET enhancement. This signal amplification mechanism cannot be explained simply based on our current results and knowledge of FRET. Yet it is possible to utilize this double-labeled donor system in various applications of FRET as a simple signal-enhancement method. 10.1093/nar/28.24.e107}, author = {Okamura, Yukio and Kondo, Satoshi and Sase, Ichiro and Suga, Takayuki and Mise, Kazuyuki and Furusawa, Iwao and Kawakami, Shigeki and Watanabe, Yuichiro }, citeulike-article-id = {2524525}, doi = {10.1093/nar/28.24.e107}, journal = {Nucl. Acids Res.}, keywords = {addons, addons-fluorescence, fret, origami}, month = {December}, number = {24}, pages = {e107+}, posted-at = {2008-03-13 09:23:46}, priority = {2}, title = {Double-labeled donor probe can enhance the signal of fluorescence resonance energy transfer (FRET) in detection of nucleic acid hybridization}, url = {http://dx.doi.org/10.1093/nar/28.24.e107}, volume = {28}, year = {2000} }

TY - JOUR ID - citeulike:2524525 L3 - citeulike-article-id:2524525 TI - Double-labeled donor probe can enhance the signal of fluorescence resonance energy transfer (FRET) in detection of nucleic acid hybridization JF - Nucl. Acids Res. VL - 28 IS - 24 SP - e107 N2 - A set of fluorescently-labeled DNA probes that hybridize with the target RNA and produce fluorescence resonance energy transfer (FRET) signals can be utilized for the detection of specific RNA. We have developed probe sets to detect and discriminate single-strand RNA molecules of plant viral genome, and sought a method to improve the FRET signals to handle in vivo applications. Consequently, we found that a double-labeled donor probe labeled with Bodipy dye yielded a remarkable increase in fluorescence intensity compared to a single-labeled donor probe used in an ordinary FRET. This double-labeled donor system can be easily applied to improve various FRET probes since the dependence upon sequence and label position in enhancement is not as strict. Furthermore this method could be applied to other nucleic acid substances, such as oligo RNA and phosphorothioate oligonucleotides (S-oligos) to enhance FRET signal. Although the double-labeled donor probes labeled with a variety of fluorophores had unexpected properties (strange UV-visible absorption spectra, decrease of intensity and decay of donor fluorescence) compared with single-labeled ones, they had no relation to FRET enhancement. This signal amplification mechanism cannot be explained simply based on our current results and knowledge of FRET. Yet it is possible to utilize this double-labeled donor system in various applications of FRET as a simple signal-enhancement method. 10.1093/nar/28.24.e107 KW - addons KW - addons-fluorescence KW - fret KW - origami AU - Okamura, Yukio AU - Kondo, Satoshi AU - Sase, Ichiro AU - Suga, Takayuki AU - Mise, Kazuyuki AU - Furusawa, Iwao AU - Kawakami, Shigeki AU - Watanabe, Yuichiro PY - 2000/12/15/ UR - http://dx.doi.org/10.1093/nar/28.24.e107 ER -