Zhenzhen Huang

Incorporating Graphene Oxide and Gold Nanoclusters: A Synergistic Catalyst with Surprisingly High Peroxidase-Like Activity Over a Broad pH Range and its Application for Cancer Cell Detection

A synergistic graphene oxide-gold nanocluster (GO-AuNC) hybrid has been constructed as an enzyme mimic that is able to show high catalytic activity over a broad pH range, especially at neutral pH. Importantly, the target-functionalized hybrid has been applied as a robust nanoprobe for selective, quantitative, and fast colorimetric detection of cancer cells.

Stimuli-responsive controlled-release system using quadruplex DNA-capped silica nanocontainers

A novel proton-fueled molecular gate-like delivery system has been constructed for controlled cargo release using i-motif quadruplex DNA as caps onto pore outlets of mesoporous silica nanoparticles. Start from simple conformation changes, the i-motif DNA cap can open and close the pore system in smart response to pH stimulus. Importantly, the opening/closing and delivery protocol is highly reversible and a partial cargo delivery can be easily controlled at will. A pH-switchable nanoreactor has also been developed to validate the potential of our system for on-demand molecular transport. This proof of concept might open the door to a new generation of carrier materials and could also provide a general route to use other functional nucleic acids/peptide nucleic acids as capping agents in the fields of versatile controlled delivery nanodevices.

DNA/Ligand/Ion-Based Ensemble for Fluorescence Turn on Detection of Cysteine and Histidine with Tunable Dynamic Range

A new type of rapid, highly sensitive, and selective fluorescence turn-on assay for detection of cysteine and histidine using a DNA/ligand/ion ensemble is developed. This assay is based on the highly specific interaction between the amino acids and the metal ions and the strong fluorescence thiazole orange (TO)/DNA probe in a competition assay format. The resulting high sensitivity and selectivity for cysteine and histidine was achieved by changing the metal ions. The system is simple in design and fast in operation and is more convenient and promising than other methods. The novel strategy eliminated the need of organic cosolvents, enzymatic reactions, separation processes, chemical modifications, and sophisticated instrumentations. The detection and discrimination process can be seen with the naked eye under a hand-held UV lamp and can be easily adapted to automated high-throughput screening. The detection limit of this method is lower than or at least comparable to previous fluorescence-based methods. The dynamic range of the sensor can be tuned simply by adjusting the concentration of metal ions. Importantly, the protocol offers high selectivity for the determination of cysteine among amino acids found in proteins and in serum samples. The assay shows great potential for practical application as a disease-associated biomarker and will be needed to satisfy the great demand of amino acid determination in fields such as food processing, biochemistry, pharmaceuticals, and clinical analysis.

A Label-Free, Quadruplex-Based Functional Molecular Beacon (LFG4-MB) for Fluorescence Turn-On Detection of DNA and Nuclease

We demonstrate a novel concept for the construction of a label-free, quadruplex-based functional molecular beacon (LFG4-MB) by using G-quadruplex motif as a substitute for Watson-Crick base pairing in the MB stem and a specific G-quadruplex binder, N-methyl mesoporphyrin IX (NMM) as a reporter. It shows high sensitivity in assays for UDG activity/inhibition and detection of DNA sequence based on the unique fluorescence increase that occurs as a result of the strong interaction between NMM and the folded quadruplex upon removal of uracil by UDG or displacement of block sequence by target DNA. The LFG4-MB is simple in design, fast in operation and could be easily transposed to other biological relevant target analysis by simply changing the recognition portion. The LFG4-MB does not require any chemical modification for DNA, which offers the advantages of simplicity and cost efficiency and obviates the possible interference with the affinity and specificity of the MB as well as the kinetic behavior of the catalysts caused by the bulky fluorescent groups. More importantly, the LFG4-MB offers great extent of freedom to tune the experimental conditions for the general applicability in bioanalysis.

Site-Specific DNA-Programmed Growth of Fluorescent and Functional Silver Nanoclusters

Precise organization of metallic nanoclusters on DNA scaffolds holds great interest for nanopatterned materials that may find uses in electronics, sensors, medicine, and many other fields. Herein, we report the site-specific growth of fluorescent silver nanoclusters by using a mismatched double-stranded DNA template. Few-atom, molecular-scale Ag clusters are found to localize at the mismatched site and the metallized DNA retains its integrity. The DNA-encapsulated nanoclusters can be utilized as functional biological probes to identify single-nucleotide polymorphisms by taking advantage of the very bright fluorescence and excellent photostability of the nanoclusters. This approach offers the possibility of constructing novel DNA-based nanomaterials and nanomechanical devices with more sophisticated functions and will be highly beneficial in future biochemical, pharmaceutical, nanomechanical, and electronic applications.

A Quadruplex-Based, Label-Free, and Real-Time Fluorescence Assay for RNase H Activity and Inhibition

We demonstrate a unique quadruplex-based fluorescence assay for sensitive, facile, real-time, and label-free detection of RNase H activity and inhibition by using a G-quadruplex formation strategy. In our approach, a RNA-DNA substrate was prepared, with the DNA strand designed as a quadruplex-forming oligomer. Upon cleavage of the RNA strand by RNase H, the released G-rich DNA strand folds into a quadruplex in the presence of monovalent ions and interacts with a specific G-quadruplex binder, N-methyl mesoporphyrin IX (NMM); this gives a dramatic increase in fluorescence and serves as a reporter of the reaction. This novel assay is simple in design, fast in operation, and is more convenient and promising than other methods. It takes less than 30 min to finish and the detection limit is much better or at least comparable to previous reports. No sophisticated experimental techniques or chemical modification for either RNA or DNA are required. The assay can be accomplished by using a common spectrophotometer and obviates possible interference with the kinetic behavior of the catalysts. Our approach offers an ideal system for high-throughput screening of enzyme inhibitors and demonstrates that the structure of the G-quadruplex can be used as a functional tool in specific fields in the future.

Toward site-specific, homogeneous and highly stable fluorescent silver nanoclusters fabrication on triplex DNA scaffolds

A new strategy to create site-specific, homogeneous, and bright silver nanoclusters (AgNCs) with high-stability was demonstrated by triplex DNA as template. By reasonable design of DNA sequence, homogeneous Ag2 cluster was obtained in the predefined position of CG.C+ site of triplex DNA. This strategy was also explored for controlled alignment of AgNCs on the DNA nanoscaffold. To the best of our knowledge, this was the first example to simultaneously answer the challenges of excellent site-specific nucleation and growth, homogeneity and stability against salt of DNA-templated AgNCs.

Artificial DNA nano-spring powered by protons.

A novel multifunctional, proton-fueled DNA nano-spring has been constructed. By incorporation of the G-quadruplex/i-motif sequence into the assembly, the nanodevice can perform springlike motions in response to changes in the environmental pH without permanent deformation. Nanosized objects/functional groups could be assembled/disassembled into this system in an addressable, contractile, and reversible manner.

Versatile Logic Devices Based on Programmable DNA-Regulated Silver-Nanocluster Signal Transducers

A DNA-encoding strategy is reported for the programmable regulation of the fluorescence properties of silver nanoclusters (AgNCs). By taking advantage of the DNA-encoding strategy, aqueous AgNCs were used as signal transducers to convert DNA inputs into fluorescence outputs for the construction of various DNA-based logic gates (AND, OR, INHIBIT, XOR, NOR, XNOR, NAND, and a sequential logic gate). Moreover, a biomolecular keypad that was capable of constructing crossword puzzles was also fabricated. These AgNC-based logic systems showed several advantages, including a simple transducer-introduction strategy, universal design, and biocompatible operation. In addition, this proof of concept opens the door to a new generation of signal transducer materials and provides a general route to versatile biomolecular logic devices for practical applications.

Poly(acrylic acid)-templated silver nanoclusters as a platform for dual fluorometric turn-on and colorimetric detection of mercury (II) ions

An easy prepared fluorescence turn-on and colorimetric dual channel probe was developed for rapid assay of Hg(2+) ions with high sensitivity and selectivity by using poly(acrylic acid)-templated silver nanoclusters (PAA-AgNCs). The PAA-AgNCs exhibited weak fluorescence, while upon the addition of Hg(2+) ions, AgNCs gives a dramatic increase in fluorescence as a result of the changes of the AgNCs states. The detection limit was estimated to be 2 nM, which is much lower than the Hg(2+) detection requirement for drinking water of U.S. Environmental Protection Agency, and the turn-on sensing mode offers additional advantage to efficiently reduce background noise. Also, a colorimetric assay of Hg(2+) ions can be realized due to the observed absorbance changes of the AgNCs. More importantly, the method was successfully applied to the determination of Hg(2+) ions in real water samples, which suggests our proposed method has a great potential of application in environmental monitoring.