We report an effective, novel self-assembled single-wall carbon nanotube (SWNT) complex with an oligonucleotide and demonstrate its feasibility in recognizing and detecting specific DNA sequences in a single step in a homogeneous solution. The key component of this complex is the hairpin-structured fluorescent oligonucleotide that allows the SWNT to function as both a "nanoscaffold" for the oligonucleotide and a "nanoquencher" of the fluorophore. Given this functionality, this carbon nanotube complex represents a new class of universal fluorescence quenchers that are substantially different from organic quenchers and should therefore have many applications in molecular engineering and biosensor development. Competitive binding of a DNA target and SWNTs with the oligonucleotide results in fluorescence signal increments relative to the fluorescence without a target as well as in marked fluorescence quenching. In contrast to the common loop-and-stem configuration of molecular beacons (MBs), this novel fluorescent oligonucleotide needs only one labeled fluorophore, yet the emission can be measured with little or no background interference. This property greatly improves the signal-to-background ratio compared with those for conventional MBs, while the DNA-binding specificity is still maintained by the MB. To test the interaction mechanisms of the fluorescent oligonucleotide with SWNTs and target DNA, thermodynamic analysis and fluorescence anisotropy measurements, respectively, were applied. Our results show that MB/SWNT probes can be an excellent platform for nucleic acid studies and molecular sensing.
Cell types, both healthy and diseased, can be classified by inventories of their cell-surface markers. Programmable analysis of multiple markers would enable clinicians to develop a comprehensive disease profile, leading to more accurate diagnosis and intervention. As a first step to accomplish this, we have designed a DNA-based device, called “Nano-Claw”. Combining the special structure-switching properties of DNA aptamers with toehold-mediated strand displacement reactions, this claw is capable of performing autonomous logic-based analysis of multiple cancer cell-surface markers and, in response, producing a diagnostic signal and/or targeted photodynamic therapy. We anticipate that this design can be widely applied in facilitating basic biomedical research, accurate disease diagnosis and effective therapy.
A highly selective copper(II) ion fluorescent sensor has been designed based on the UV-visible absorption of a spiropyran derivative coupled with the use of a metal porphyrin operative on the fluorescence inner filter effect. Spiropyrans, which combine the characteristics of metal binding and signal transduction, have been widely utilized in cationic ion recognition by UV-visible spectroscopy. In the present work, the viability of converting the absorption signal of the spiropyran molecule into a fluorescence signal was explored. On account of overlap of the absorption band of the spiropyran (lambda(abs) = 547 nm) in the presence of copper ion with the Q-band of an added fluorophore, zinc meso-tetraphenylporphyrin (lambda(abs) = 556 nm), the effective light absorbed by the porphyrin and concomitantly the emitted light intensity vary as a result of varying absorption of the spiropyran via fluorescence inner filter effect. The metal binding characteristic of the spiropyran presents an excellent selectivity for copper ion in comparison with several other heavy or transition metal ions. Since the changes in the absorbance of the absorber translate into exponential changes in fluorescence of the fluorophore, the novelty of the present device is that the analytical signal is more sensitive over that of the absorptiometry or that of the fluorometry using one single dye. To realize a practical fluorescent sensor, both the absorber and fluorophore were immobilized in a plasticized poly(vinyl chloride) membrane, and the sensing characteristics of the membrane for copper ion were investigated. The sensor is useful for measuring Cu2+ at concentrations ranging from 7.5 x 10(-7) to 3.6 x 10(-5) M with a detection limit of 1.5 x 10(-7) M. The sensor is chemically reversible, the fluorescence was switched off by immersing the membrane in copper ion solution and switched on by washing it with EDTA solution.
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