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A novel coumarin-based fluorescent probe, P-CM, for quantitative detection of nitroxyl (HNO) was developed. P-CM exhibits a selective response to HNO over other biological reductants and was also applied for quantitative detection of HNO in bovine serum with satisfactory results. [ABSTRACT FROM AUTHOR]

A highly selective electrochemical biosensor for the ultrasensitive detection of Hg2+in aqueous solution has been developed based on the strong and specific binding of Hg2+by two DNA thymine bases (T–Hg2+–T) and the use of AuNP-functionalized reporter DNA to achieve signal amplification. [ABSTRACT FROM AUTHOR]

Carboxylesterase (CE) is crucial in metabolizing ester-containing biomolecules and is particularly significant in liver metabolic diseases. Herein, we present the first activatable NIRF/PA dual-mode imaging probe QHD-CE for detection of CE in vitro and in vivo. QHD-CE displays excellent sensitivity and selectivity for CE with a high reaction efficiency (∼90 min). By utilizing QHD-CE, the dynamic changes of CE in drug-induced liver injury and diabetic mice models were monitored. [ABSTRACT FROM AUTHOR]

Recently, supramolecular chemistry with its unique properties has received considerable attention in many fields. Supramolecular fluorescent systems constructed on the basis of macrocyclic hosts are not only effective in overcoming the limitations of imaging and diagnostic reagents, but also in enhancing their performances. This paper summarizes the recent advances in supramolecular fluorescent systems based on host–guest interactions and their application in bioimaging and therapy as well as the challenges and prospects in developing novel supramolecular fluorescent systems. [ABSTRACT FROM AUTHOR]

Herein, we report a new click-activated prodrug, CA4V, and a bioorthogonal nanoreactor, CA4V/ZIF-90@TzCOF@Apt, which consists of a ZIF-90 core, tetrazine-based covalent organic framework (COF) shells and an aptamer polymer coating. When targeting a tumor, the acid-causing collapse of ZIF-90 initiates a nanoconfined bioorthogonal reaction in defined COF cages, which boosts the click efficiency of CA4V activation and therapeutic effects in vivo. [ABSTRACT FROM AUTHOR]

Stable and sensitive ctDNA biosensing in complex biological fluid is highly important but still remains a challenge. Herein, we develop a spherical nucleic acid reporter-based cascade CRISPR/Cas12a amplifier with improved stability and sensitivity (5 orders of magnitude). [ABSTRACT FROM AUTHOR]

We report here an activatable chemiluminescent probe CL-O3 for the high-contrast imaging of O3in vivo. CL-O3 exhibited a high selectivity toward O3 and was able to evaluate the degree of inflammation in mice by detecting endogenous O3 levels in acute inflamed mice. [ABSTRACT FROM AUTHOR]

In situ monitoring of the location and transportation of bioactive molecules is essential for deciphering diverse biological events in the field of biomedicine. In addition, obtaining the in situ information of lesions will provide a clear perspective for surgeons to perform precise resection in clinical surgery. Notably, delivering drugs or operating photodynamic therapy/photothermal therapy in situ by labeling the lesion regions of interest can improve treatment and reduce side effects in vivo. In various advanced imaging and therapy modalities, optical theranostic agents based on organic small molecules can be conveniently modified as needed and can be easily internalized into cells/lesions in a non-invasive manner, which are prerequisites for in situ bioimaging and precision treatment. In this tutorial review, we first summarize the in situ molecular immobilization strategies to retain small-molecule agents inside cells/lesions to prevent their diffusion in living organisms. Emphasis will be focused on introducing the application of these strategies for in situ imaging of biomolecules and precision treatment, particularly pertaining to why targeting therapy in situ is required. [ABSTRACT FROM AUTHOR]

The sensitive and accurate detection of mature miRNA without the signal interference by pre-miRNAs is highly important. Herein, a size-selective DNA nanocage-based activatable CRISPR/Cas12a system was developed to achieve this goal. [ABSTRACT FROM AUTHOR]

Natural living systems are driven by delicate protein networks whose functions are precisely controlled by many parameters, such as number, distance, orientation, and position. Focusing on regulation rather than just imitation, the construction of artificial protein networks is important in many research areas, including biomedicine, synthetic biology and chemical biology. DNA origami, sophisticated nanostructures with rational design, can offer predictable, programmable, and addressable scaffolds for protein assembly with nanometer precision. Recently, many interdisciplinary efforts have achieved the precise construction of DNA origami-based protein networks, and their emerging application in many areas. To inspire more fantastic research and applications, herein we highlight the applicability and potentiality of DNA origami-based protein networks. After a brief introduction to the development and features of DNA origami, some important factors for the precise construction of DNA origami-based protein networks are discussed, including protein–DNA conjugation methods, networks with different patterns and the controllable parameters in the networks. The discussion then focuses on the emerging application of DNA origami-based protein networks in several areas, including enzymatic reaction regulation, sensing, bionics, biophysics, and biomedicine. Finally, current challenges and opportunities in this research field are discussed. [ABSTRACT FROM AUTHOR]

The in situ and real-time supervision of reactive oxygen species (ROS) generated during photodynamic therapy (PDT) is of great significance for lessening nonspecific damage and guiding personalized therapy. However, photosensitizers frequently fail to deliver successful treatment accompanying the ROS-related imaging signals produced, impeding simple treatment outcome predictions and therapeutic schedule adjustments. Here, we report a two-photon fluorescence self-reporting strategy for the in situ and real-time monitoring of treatment response via a novel black phosphorus-based two-photon nanoprobe (TPBP). TPBP effectively generated singlet oxygen (1O2) under near-infrared laser irradiation for PDT, and 1O2 stimulated a two-photon molecule to emit fluorescence signals for feedback of 1O2 generation, which facilitated the regulation of treatment parameters to achieve precise and personalized medicine in deep tissue. [ABSTRACT FROM AUTHOR]

A FRET-based probe for mapping the fluctuation of ONOO− in cisplatin-induced acute kidney injury was constructed. It exhibits ratiometric near infrared fluorescence and a dramatic decrease of its peak absorbance at 719 nm upon addition of ONOO− that is converted into remarkable signal changes in fluorescence and photoacoustic images respectively. [ABSTRACT FROM AUTHOR]

The enzymatic-assisted signal amplification of DNA sensors is rarely applied in living cells due to the difficulties in protein delivery. In this study, we have proposed a biomineralization-based DNA nanoprobe to transport nucleases and DNA sensors for enzyme-assisted imaging of microRNA in living cells. [ABSTRACT FROM AUTHOR]

Herein, we report a pH stimulus-disaggregated BODIPY sensitizer (PTS) with low background-toxicity for achieving activated photodynamic/photothermal tumor therapy. Both the photodynamic and photothermal properties of PTS can be activated under acidic conditions, and PTS exhibits excellent antitumor properties, which is revealed by both in vitro and in vivo tests. [ABSTRACT FROM AUTHOR]

By assembling DNAzyme on DNA nanowires through DNA hybridization, we have developed a novel accelerated DNAzyme-based fluorescent nanoprobe for fast, sensitive and selective detection of miRNA. Moreover, the strategy was successfully applied for in situ imaging of miRNA-21 in different cell lines. [ABSTRACT FROM AUTHOR]

We report here a single-benzene based fluorescent framework, amino-terephthalonitrile, denoted SB-Fluor. This scaffold displays versatile emission wavelength tunability via structure modification, covering the full visible light spectrum, both in the solution and solid state. Moreover, one molecule, SBF3, exhibits polymorphism-dependent reversible mechanochromic luminescence. [ABSTRACT FROM AUTHOR]

A highly selective lysosome-targeting NIR fluorescent probe (Lyso-SiR-2S) for HOCl was constructed based on Si-rhodamine B spirodithiolactone. This probe was very effectively employed to sense HOCl produced in various living cells and to visualize fluctuations of endogenous HOCl resulting from GEN-induced acute kidney injury in vivo for the first time. [ABSTRACT FROM AUTHOR]

In this work, we report the first bioluminescent probe BP-PS for detecting H2Sn with high specificity and sensitivity. Owing to the bioluminescence imaging without requiring an excitation light source, tissue autofluorescence is eliminated and BP-PS shows a high signal-to-noise ratio. Moreover, BP-PS was successfully utilized to visualize endogenous H2Sn in live cells and a murine model of bacterial infection. [ABSTRACT FROM AUTHOR]

Precision diagnosis and effective treatment are the cores of early cancer therapy. Here, for the first time, we report a hybridization chain reaction-based nanoprobe for selective and sensitive cancer cell recognition and amplified photodynamic therapy. [ABSTRACT FROM AUTHOR]

A bioluminescent probe, BP-HNO, which exhibits a turn-on response to nitroxyl with high sensitivity and selectivity, is reported for the first time in this work. BP-HNO is free from the interference of biological autofluorescence to afford a high signal-to-noise ratio for bioimaging, and was successfully applied to imaging nitroxyl in live cells and mice. [ABSTRACT FROM AUTHOR]

Abnormal enzymatic activities are directly related to the development of cancers. Identifying the location and expression levels of these enzymes in live cancer cells have considerable importance in early-stage cancer diagnoses and monitoring the efficacy of therapies. Small-molecule fluorescent probes have become a powerful tool for the detection and imaging of enzymatic activities in biological systems by virtue of their higher sensitivity, nondestructive fast analysis, and real-time detection abilities. Moreover, due to their structural tailorability, numerous small-molecule enzymatic fluorescent probes have been developed to meet various demands involving real-time tracking and visualizing different enzymes in live cancer cells or in vivo. In this review, we provide an overview of recent advances in small-molecule enzymatic fluorescent probes mainly during the past decade, including the design strategies and applications for various enzymes in live cancer cells. We also highlight the challenges and opportunities in this rapidly developing field of small-molecule fluorescent probes for interventional surgical imaging, as well as cancer diagnosis and therapy. [ABSTRACT FROM AUTHOR]

The development of selective and sensitive sensing systems for Cr(vi) anions (CrO42−/Cr2O72−) has been a growing interest because Cr(vi) anions are potent carcinogenic and mutagenic ions that have become serious potential threats to public health and safety. Herein, the selective and highly sensitive detection of toxic Cr(vi) anions in aqueous solution has been achieved by virtue of nanoscale zeolitic imidazole framework-90 (NZIF-90). NZIF-90 is the first individual MOF-based material which can selectively sense Cr(vi) anions from Fe3+ cations, and it also has the lowest detection limits among the reported fluorescent MOF-based probes for Cr(vi) anions: 1.41 μM (0.16 ppm) for CrO42− and 1.78 μM (0.38 ppm) for Cr2O72−. The detailed sensing mechanism study uncovered that the excellent sensing performance of NZIF-90 for Cr(vi) anions is based on the synergistic effect of the competitive energy absorption process and the electrostatic interaction between NZIF-90 and Cr(vi) anions. Moreover, as for a further potential application of this probe, the Förster resonance energy transfer (FRET)-based dual-excitation ratiometric fluorescent detection of Cr(vi) anions was also realized by Rhodamine B (RhB) encapsulated into NZIF-90 (RhB@NZIF-90). [ABSTRACT FROM AUTHOR]

Herein, we developed a strategy to attack the cancer cell defense system against reactive oxygen species to improve photodynamic therapy efficacy with a Ce6@MSN@MTH1 siRNA nanosystem, which was demonstrated to improve cellular sensitivity to reactive oxygen species through suppressing MTH1 protein in cancer cells. [ABSTRACT FROM AUTHOR]

Oxidative stress, a disturbance in the balance between oxidant/antioxidant ratios, is associated with cancer, aging, inflammation, neurodegenerative diseases and other conditions. γ-Glutamyltranspeptidase (GGT) is a redox-related enzyme that plays a key role in mitigating the effects of oxidative stress by maintaining cellular glutathione (GSH) metabolism and homeostasis. Therefore, oxidative stress will upregulate the intracellular GGT level. To better understand the major pathophysiological resist mechanism to oxidative injury in mediating many disease states, we designed and synthesized a novel two-photon (TP) fluorescent turn-on probe, Np-Glu, for GGT detection and bioimaging. Under the optimized conditions, Np-Glu exhibited remarkable fluorescence enhancement (150-fold), good selectivity and high sensitivity (LOD is 0.033 U L−1), with a wide linear concentration range (0–50 U L−1). More importantly, the probe Np-Glu was successfully applied in one-photon and TP fluorescence imaging of GGT activity in an oxidative stress model in living cells and tissues, suggesting Np-Glu as an ideal indicator for clinical and biological samples. [ABSTRACT FROM AUTHOR]

A promising two-photon fluorescent probe MNAH for detecting 1O2 during the PDT process in mitochondria was proposed for the first time. MNAH was successfully applied for two-photon imaging of 1O2 in living cells and tissues during the PDT process with deep-tissue imaging depth. MNAH can be a powerful molecular tool for studying 1O2 generation in mitochondria during the PDT process. [ABSTRACT FROM AUTHOR]

Cellular self-regulation of reactive oxygen species (ROS) stress via antioxidant repair plays an important role in maintaining the redox balance. The redox balance between reducing and oxidizing species within cells is significant in the regulation of a signal pathway and is achieved by a series of elaborate mechanisms. In this work, we employed our previously reported D–π-A-structured naphthalene–BODIPY TBET platform to design an efficient two-photon fluorescent probe for dynamic monitoring of superoxide anion oxidative stress and the GSH reducing repair process. The probe displayed high energy transfer efficiency (91.4%), large pseudo-Stokes shifts upon one-photon excitation, and red fluorescence emission (λem = 596 nm), which is highly desirable for bioimaging applications. The probe exhibits reversibility, rapid response, good photostability, high selectivity and sensitivity for the superoxide anion and GSH. More importantly, the probe was successfully applied for visualizing the redox changes in living cells and tissues. [ABSTRACT FROM AUTHOR]

Formaldehyde (FA) plays an important role in living systems as a reactive carbonyl species (RCS). An abnormal degree of FA is known to induce neurodegeneration, cognitive decrease and memory loss owing to the formation of strong cross-link DNA and protein and other molecules. The development of efficient methods for biological FA detection is of great biomedical importance. Although a few one-photon FA fluorescent probes have been reported for imaging in living cells, probes excited by two photons are more suitable for bio-imaging due to their low background fluorescence, less photobleaching, and deep penetration depth. In this study, a two-photon fluorescent probe FATP1 for FA detection and bio-imaging in living cells and tissues was reported. The detection is based on the 2-aza-Cope sigmatropic rearrangement followed by elimination to release the fluorophore, resulting in both one- and two-photon excited fluorescence increase. The probe FATP1 showed a high sensitivity to FA with a detection limit of 0.2 μM. Moreover, FATP1 enabled the two-photon bio-imaging of FA in live HEK-293 cells and tissues with tissue-imaging depths of 40–170 μm. Furthermore, FATP1 could be applied for the monitoring of endogenous FA in live MCF-7 cells, presaging its practical applications in biological systems. [ABSTRACT FROM AUTHOR]

The combination of nanostructures with biomolecules leading to the generation of functional nanosystems holds great promise for biotechnological and biomedical applications. As a naturally occurring biomacromolecule, DNA exhibits excellent biocompatibility and programmability. Also, scalable synthesis can be readily realized through automated instruments. Such unique properties, together with Watson–Crick base-pairing interactions, make DNA a particularly promising candidate to be used as a building block material for a wide variety of nanostructures. In the past few decades, various DNA nanostructures have been developed, including one-, two- and three-dimensional nanomaterials. Aptamers are single-stranded DNA or RNA molecules selected by Systematic Evolution of Ligands by Exponential Enrichment (SELEX), with specific recognition abilities to their targets. Therefore, integrating aptamers into DNA nanostructures results in powerful tools for biosensing and bioimaging applications. Furthermore, owing to their high loading capability, aptamer-modified DNA nanostructures have also been altered to play the role of drug nanocarriers for in vivo applications and targeted cancer therapy. In this review, we summarize recent progress in the design of aptamers and related DNA molecule-integrated DNA nanostructures as well as their applications in biosensing, bioimaging and cancer therapy. To begin with, we first introduce the SELEX technology. Subsequently, the methodologies for the preparation of aptamer-integrated DNA nanostructures are presented. Then, we highlight their applications in biosensing and bioimaging for various targets, as well as targeted cancer therapy applications. Finally, we discuss several challenges and further opportunities in this emerging field. [ABSTRACT FROM AUTHOR]

Cell-surface fluorescent probes are effective tools in cell biology and engineering. Here, we for the first time report a diacyllipid–aptamer conjugate-based fluorescent probe which could anchor on cell membrane for real-time tracking of potassium ions in the cell microenvironment. [ABSTRACT FROM AUTHOR]

A FRET-based two-photon fluorescent probe, P-Np-Rhod, which exhibited a fast and high selective ratiometric response to nitroxyl, was first proposed. P-Np-Rhod was successfully applied to two-photon dual-channel imaging of nitroxyl in living cells and tissues with less cross-talk between channels and satisfactory deep-tissue imaging depth. [ABSTRACT FROM AUTHOR]

Lysosomes are acidic organelles (approximately pH 4.5–5.5) and tracking the changes in lysosomal pH is of great biological importance. To address this issue, quite a few of fluorescent probes have been developed. However, few of these probes can realize the tracking of dynamic changes in lysosomal pH. Herein, we report a new lysosome-targeted ratiometric fluorescent probe (FR-Lys) by hybridizing morpholine with a xanthane derivative and an o-hydroxy benzoxazole group. In this probe, the morpholine group serves as a targeting unit for lysosome, the xanthane derivative exhibits a pH-modulated open/close reaction of the spirocycle, while the o-hydroxy benzoxazole moiety shows a pH modulated excited-state intramolecular proton transfer (ESIPT) process. Such a design affords the probe a ratiometric fluorescence response towards pH with pH values ranging from 4.0 to 6.3. The response of the probe to pH was fast and reversible with high selectivity. Moreover, this probe possesses further advantages such as easy synthesis, high photostability and low cytotoxicity. These features are favorable for tracking dynamic pH changes in biosystems. It was then applied for dynamic imaging pH changes in lysosomes with satisfactory results. [ABSTRACT FROM AUTHOR]

By employing DNAzyme as a recognition group and amplifier, and DNA-stabilized silver nanoclusters (DNA/AgNCs) as signal reporters, we reported for the first time a label-free catalytic and molecular beacon as an amplified biosensing platform for highly selective detection of cofactors such as Pb2+ and l-histidine. [ABSTRACT FROM AUTHOR]

Taking advantage of the super-quenching effect of the cationic perylene derivative on adjacent fluorophores, we for the first time reported a DNAzyme–perylene complex-based strategy for constructing fluorescence catalytic biosensors with improved sensitivity. [ABSTRACT FROM AUTHOR]

The use of a nanoscale DNA–Au dendrimer as a signal amplifier was proposed for the universal design of functional DNA-based ultra-sensitive SERS biosensors. This novel design combines the high specificity of functional DNA with the high sensitivity of surface-enhanced Raman scattering (SERS) spectroscopy, resulting in sensitivity superior to that of previously reported sensors. [ABSTRACT FROM AUTHOR]