Chunyang Lei

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Verified

Chunyang verified their affiliation via an institutional email.

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• Ph.D.
• Professor (Full) at Hunan University

Viral proteases play a crucial role in viral infection and are regarded as promising targets for antiviral drug development. Consequently, biosensing methods that target viral proteases have contributed to the study of virus-related diseases. This work presents a ratiometric electrochemical sensor that enables highly sensitive detection of viral pr...

Natural protein–protein communications, such as those between transcription factors (TFs) and RNA polymerases/ribosomes, underpin cell‐free biosensing systems operating on the transcription/translation (TXTL) paradigm. However, their deployment in field analysis is hampered by the delayed response (hour‐level) and the complex composition of in vitr...

Proteins capable of switching between distinct active states in response to biochemical cues are ideal for sensing and controlling biological processes. Activatable CRISPR‐Cas systems are significant in precise genetic manipulation and sensitive molecular diagnostics, yet directly controlling Cas protein function remains challenging. Herein, we exp...

Fluorogenic RNA aptamers have revolutionized the visualization of RNAs within complex cellular processes. A representative category of them employs the derivatives of green fluorescent protein chromophore, 4‐hydroxybenzlidene imidazolinone (HBI), as chromophores. However, the structural homogeneity of their chromophoric backbones causes severe cros...

Fluorogenic RNA aptamers have revolutionized the visualization of RNAs within complex cellular processes. A representative category of them employs the derivatives of green fluorescent protein chromophore, 4‐hydroxybenzlidene imidazolinone (HBI), as chromophores. However, the structural homogeneity of their chromophoric backbones causes severe cros...

G-quadruplexes (G4s) are non-canonical nucleic acid secondary structures that play a crucial role in regulating essential cellular processes such as replication, transcription, and translation. The formation of G4s is dynamically...

Engineered living materials (ELMs) harness engineered cells to fabricate functional materials with lifelike characteristics, offering unparalleled potential across various fields. Nonetheless, the deployment of ELM‐based biosensors beyond laboratory settings remains challenging. Herin, ELMs are explored as field‐deployable biosensing laboratories o...

The precise target recognition and nuclease-mediated effective signal amplification capacities of CRISPR-Cas systems have attracted considerable research interest within the biosensing field. Guided by insights into their structural and biochemical mechanisms, researchers have endeavored to engineer the key biocomponents of CRISPR-Cas systems with...

Natural protein–protein communications, such as those between transcription factors (TFs) and RNA polymerases/ribosomes, underpin cell‐free biosensing systems operating on the transcription/translation (TXTL) paradigm. However, their deployment in field analysis is hampered by the delayed response (hour‐level) and the complex composition of in vitr...

In vitro RNA synthesis technologies are crucial in developing therapeutic RNA drugs, such as mRNA vaccines and RNA interference (RNAi) therapies. Enzymatic RNA synthesis, recognized for its sustainability and efficiency, enables the production of extensive RNA sequences under mild conditions. Among the enzymes utilized, T7 RNA polymerase is disting...

Proteins capable of switching between distinct active states in response to biochemical cues are ideal for sensing and controlling biological processes. Activatable CRISPR‐Cas systems are significant in precise genetic manipulation and sensitive molecular diagnostics, yet directly controlling Cas protein function remains challenging. Herein, we exp...

Programmable and precise regulation of genetic information is crucial in bioengineering and biomedicine; however, it remains challenging to implement this objective. Here we deployed DNA-functionalized MXenes as a smart delivery system for spatiotemporally controllable genome editing. The MXene nanovehicles rationally integrated photothermal effect...

The CRISPR/Cas12a system has been repurposed as a versatile nuclei acid bio-imaging tool, but its utility in sensing non-nucleic acid analytes in living cells has been less exploited. Herein, we demonstrated the ability of Mn2+ to accelerate cleavage kinetics of Cas12a and deployed for live-cell Mn2+ sensing by leveraging the accelerated trans-clea...

G-quadruplex (G4) is a noncanonical nucleic acid secondary structure that has implications for various physiological and pathological processes and is thus essential to exploring new approaches to G4 detection in live cells. However, the deficiency of molecular imaging tools makes it challenging to visualize the G4 in ex vivo tissue samples. In thi...

The efficient and robust signal reporting ability of CRISPR-Cas system exhibits huge value in biosensing, but its applicability for non-nucleic acid analyte detection relies on the coupling of additional recognition modules. To address this limitation, we described a switchable Cas12a and exploited it for CRISPR-based direct analysis of histone dea...

With increasing concerns about health, safety, and environmental issues, the development of sensitive, reliable and efficient biosensing technologies for the early detection of disease biomarkers and toxic chemicals is highly desirable. Electrochemical techniques have the advantages of low cost, high sensitivity and easy miniaturization. Cell-free...

The CRISPR-Cas system has been repurposed as a powerful live-cell imaging tool, but its utility is limited to genomic loci and mRNA imaging in living cells. Here, we demonstrated the potential of the CRISPR-Cas system as a generalizable live-cell biosensing tool by extending its applicability to monitor diverse intracellular biomolecules. In this w...

Herein, we describe a CRISPR-Cas12a sensing platform activated by a DNA ligation reaction for the sensitive detection of non-nucleic acid targets, including NAD+, ATP and polynucleotide kinase (PNK). In this design, the DNA ligation reaction triggered by these biomolecules generates DNA duplexes, which can activate the nuclease activity of Cas12a t...

Cell-free synthetic biology provides a promising strategy for developing high-performance biosensors by integrating with advanced testing technologies. However, the combination of synthetic biology with electrochemical testing techniques is still underdeveloped. Here, we proposed an electrochemical biosensor for the label-free and ultrasensitive de...

Simple and visual quantitative detection of foodborne pathogens can effectively reduce the outbreaks of foodborne diseases. Herein, we developed a simple and sensitive quantum dot (QD)-based paper device for visual and quantitative detection of Escherichia coli (E. coli) O157:H7 based on immunomagnetic separation and nanoparticle dissolution-trigge...

Sensitive and facile detection of biomarkers is essential for early diagnosis and treatment of diseases. To this end, we here proposed a colorimetric protease assay by the modular combination of proteolysis-responsive transcription and spherical nucleic acids (SNAs). In this assay, target protease-mediated proteolysis triggers the synthesis of RNAs...

Artificial nucleic acid circuits with precisely controllable dynamic and function have shown great promise in biosensing, but their utility in molecular diagnostics is still restrained by the inability to process genomic DNA directly and moderate sensitivity. To address this limitation, we present a CRISPR-Cas–powered catalytic nucleic acid circuit...

The efficient signal amplification capacity of several class 2 CRISPR-Cas systems with trans-cleavage activity has exhibited great value in molecular diagnostics, but its potential application for non-nucleic-acid targets is yet underdeveloped. Here, we deploy CRISPR-Cas system for the ultrasensitive detection of protease biomarkers by the coupling...

Proteases play crucial roles in the malignant progression of tumor and thus have been regarded as biomarkers for many cancers. Although protease assays such as immunoassays and fluorogenic substrate probes have been developed, it remains challenging for them to give consideration to both sensitivity and accuracy. Here, we describe a proteolysis-res...

Lipid transferase-catalyzed protein lipidation plays critical roles in many physiological processes and it has been an increasingly attractive therapeutic target from cancer to neurodegeneration, while sensitive detection of lipid transferase activity in biological samples remains challenging. Here, we presented an AuNP-based colorimetric method wi...

CRISPR-based diagnostics (CRISPR-Dx) has shown great promise in molecular diagnostics, but its utility in the sensing of microRNA (miRNA) biomarkers is limited by sensitivity, cost and robustness. Here, we describe a CRISPR-Dx method for the sensitive and cost-effective detection of miRNAs by rationally integrating CRISPR-Cas12a with DNA circuits....

Histone demethylases (HDMs) are vital players in epigenetic regulation and important targets in cancer treatment, while the effective molecular tools for analyzing HDMs activity are still limited. Interestingly, we found that the process of Ag+-triggered oxidation of O-phenylenediamine (OPD) to 2,3-diaminophenazine (OPDox) could be efficiently inhi...

Foodborne diseases caused by pathogens are great threats to human health. Rapid detection of foodborne pathogens at an early stage is imperative for preventing the outbreak of diseases. The identification and detection of foodborne pathogens by biosensors have attracted great attention due to their high sensitivity, nearly real-time quantification...

Post-translational modifications (PTMs) refer to the chemical modifications of proteins coordinated by PTM enzymes, and play a key role in numerous physiological and pathological processes. Herein, chimeric peptide-functionalized titanium carbide MXenes (Pep-Ti3C2) were devised for the activity assay of PTM enzymes by the integration with carboxype...

In cellular microenvironment, numerous biomolecules involved in physiological and pathological processes. To in-depth and comprehensive understanding of their roles at the molecular level, there is still a lack of detecting techniques for in situ tracking these biomolecules in local environment. Herein, we engineered the membrane-inserting peptide...

Signal amplification is an effective way to achieve sensitive analysis of biomarkers, exhibiting great promise in biomedical research and clinical diagnosis. Inspired by the transcription process, we here present a versatile strategy that enables to effectively amplify proteolysis into nucleic acid signal outputs in homogeneous system. In this stra...

The efficient protein delivery into the target cell is highly desirable for protein therapeutics. Current approaches for protein delivery commonly suffer from low-loading protein capacity, poor specificity for target cells, and invisible protein release. Herein we report a [email protected] nano-dumpling (ND) system as an intracellular protein deli...

The system of clustered regularly interspaced short palindromic repeats(CRISPR) and CRISPR-associated endonucleases(Cas) have been widely used in gene editing, disease treatment, molecular diagnosis and chromosome imaging. On account of the programmable target recognition of CRISPR-Cas system and the specific targeting function toward RNA of type V...

A near-infrared (NIR) fluorescent nanoprobe that enables to circumvent the interference of background absorption and fluorescence in whole blood, was developed for the direct sensing of blood glucose. Here, NIR fluorescent protein (iRFP) and glucose oxidase (GOx) were collectively deployed as the templates for the biomineralization of Mn²⁺ to prepa...

Fluorescent proteins (FPs) are powerful and fundamental tools in bio-analytical research, ranging from tracking biological molecules to monitoring dynamic processes in living cells. Currently, great efforts have been made on structurally engineering of FPs with new fluorescent signal-switch mechanisms via rational or semi-rational design. In this r...

A click-type protein-DNA conjugation, named as MnDDC (Mn²⁺-activated DCV-DNA conjunction), is presented, where DCV (rep protein of duck circovirus) and its target DNA work as the modular blocks to rapidly and effectively generate Mn²⁺-dependent and site-specific protein-DNA linkage. On the basis of MnDCC, a fluorescent Mn²⁺ biosensor composed of DC...

The visualization of the long noncoding RNA of prostate cancer gene 3 (lncRNA PCA3), a specific biomarker for androgen receptor-positive prostate cancer, in living cells not only directly reflects the gene expression and localization, but also offers a better insight into its roles in the pathological processes. Here, we loaded an entropy-driven RN...

A modular strategy for the construction of cell surface DNA sensors is presented by Z. Nie, C. Lei, and co‐workers in their Communication (DOI: 10.1002/anie.201901320). By engineering DNA motifs and synthetic cofactors, they developed cell surface DNA sensors for ratiometric imaging of the extrusion of endogenous signaling molecules, including sulf...

Eine modulare Strategie zur Konstruktion von Zelloberflächen‐DNA‐Sensoren wird von Z. Nie, C. Lei et al. in ihrer Zuschrift (DOI: 10.1002/ange.201901320) vorgestellt. Durch Manipulation von DNA‐Motiven und synthetischen Cofaktoren wurden Zelloberflächen‐DNA‐Sensoren für die ratiometrische Abbildung der Freisetzung endogener Signalmoleküle, inkl. Sc...

The comprehensive understanding of the mechanisms underlying the interaction of cells with their membrane microenvironment is of great value for fundamental biological research; however, tracking biomolecules on cell surfaces with high temporal and spatial resolution remains a challenge. Herein, a modular strategy is presented for the construction...

The comprehensive understanding of the mechanisms underlying the interaction of cells with their membrane microenvironment is of great value for fundamental biological research; however, tracking biomolecules on cell surfaces with high temporal and spatial resolution remains a challenge. Herein, we presented a modular strategy for the construction...

A semisynthetic fluorescent protein assembly-based FRET probe (sFPAP) was proposed for cell membrane protease function assay. Here, the probe was anchored on living cell membrane through membrane inserting peptide (MIP),...

Acquiring the multilayer information of diverse biomarkers with different spatial distributions at cellular level is crucial for monitoring the progression of cancers. Herein, a dual signal-tagged chimeric DNA-functionalized titanium carbide MXenes nanoprobe (dcDNA-Ti3C2) that responds to biomarkers with different cellular locations from plasma mem...

Precise and dynamic imaging of extracellular pH is one of crucial yet challenging tasks for studying cell physiological and pathological processes. Here we construct a DNA tweezer to dynamically monitor pH change of cellular microenvironment. The DNA tweezer contains three key elements: a 3 strands ssDNA-frame labelled with cholesterol to anchor on...

As an emerging two-dimensional material, MXenes have attracted much attention due to their unique physicochemical properties, but their application in biosensing has lagged far behind because of the poor salt tolerance. Herein, a titanium carbide MXene quantum dots (Ti3C2 QDs)-based fluorescent probe for alkaline phosphatase (ALP) activity assay an...

A charge designable and tunable green fluorescent protein (GFP)-based protein delivery strategy was proposed. The acquired His 29 GFP selectively permeates cell membrane at target pH 6.5 and escapes from endosome efficiently....

As one of emerging inorganic graphene analogues, two-dimensional titanium carbide (Ti3C2) nanosheets have attracted extensive attention in recent years due to their remarkably structural and electronic properties. Herein, a sensitive and selective nanoprobe for fluorescent probing the phospholipase D activity was developed based on ultrathin Ti3C2...

As an emerging category of fluorophores, nucleic acid-stabilized silver nanoclusters (DNA/AgNCs) have attracted a range of interests and have been widely applied for interdisciplinary research. In this work, we constructed a novel DNA/AgNCs probe for cell apoptosis detection and imaging based on enzyme-polymerized polyadenylic acid (poly-dA) DNA ch...

Transpeptidation of surface proteins catalyzed by the transpeptidase sortase plays a critical role in the infection process of gram-positive pathogen, and probing sortase activity and screening its inhibitors are of great significance to fundamental biological research and pharmaceutical development, especially novel anti-virulence drug design. Her...

Herein, a light-up non-thiolated aptasensor was developed for low-mass soluble amyloid-β40 oligomers (LS-Aβ40-O). Au nanoparticle (AuNP) was employed as a colorimetric probe, and the non-thiolated aptamer (Apt) was adsorbed on AuNP surfaces, acting as a binding element for LS-Aβ40-O. The aggregation of AuNPs was induced when Apt-modified AuNPs (Apt...

A serious impetus always exists to exploit new methods to enrich the prospect of nanomaterials. Here, we report electrochemical conversion (ECC) of magnetic nanoparticles (MNPs) to electroactive Prussian blue (PB) analogues accompanying with multiple interfacial effects and its exploitation for novel label self-sacrificial biosensing strategy. The...

A versatile bio-logic system is proposed using a surface charge tunable fluorescent protein (H 39 GFP) as a generic processor. On the basis of cell penetration of H 39 GFP triggered by H ⁺ and...

Red fluorescent proteins (RFPs) have emerged as valuable biological markers for biomolecule imaging in living systems. Developing artificial fluorogenic systems that mimic RFPs remains an unmet challenge. Here, we describe the design and synthesis of six new chromophores analogous to the chromophores in RFPs. We demonstrate, for the first time, tha...

The development of rapid and sensitive detection of foodborne pathogens could effectively prevent or reduce the outbreaks of foodborne diseases. Herein, a simple, rapid, and sensitive detection of E. coli O157:H7 was developed based on antimicrobial peptide functionalized magnetic nanoparticles (MNP-AMP) and urease-catalyzed signal amplification. I...

Rapid and sensitive detection of foodborne pathogens can effectively prevent or reduce the outbreaks of foodborne diseases. In this study, an antimicrobial peptide (AMP)-based colorimetric bioassay was developed for rapid and sensitive detection of Escherichia coli O157:H7. AMP was conjugated with horseradish peroxidase (AMP–HRP) to develop a signa...

Facile regulation and enhancing of the performance of bio-immobilization materials is a key factor for their applications for biosensing, biocatalysis, bioreactor, and so on. Here, we propose a method of combined polymerizations of chemical oxidation and metal organic coordination to develop enhanced bio-immobilization matrices for high performance...

In this study, a novel label-free colorimetric assay for the detection of lipopolysaccharides (LPSs) was developed based on a LPS-binding peptide (LBP) and unmodified gold nanoparticles (AuNPs). Cationic LBP probes with a C-terminal cysteine thiol are used to anchor the surface of unmodified AuNPs with anionic surface charge and cause the aggregati...

Thiols play a curial role in various physiological functions, and the discrimination of thiols is a significant but difficult issue. Herein, we presented a new strategy for strengthening the discrimination of thiols by a facile colorimetric sensor array composed of a series of urease-metal ion pairs. The proposed sensor array was fabricated based o...

DNA-related enzymes, including nucleases and DNA modifying enzymes, play key roles in DNA metabolism. Herein, a versatile platform using supercharged green fluorescence protein (ScGFP) as a recognition and signal output element was developed for homogeneous, turn-on, rapid, and sensitive detection of various DNA-related enzymes, including endonucle...

DNA methylation, catalyzed by DNA methyltransferases (MTases), is a key component of genetic regulation, and DNA MTases have been regarded as potential targets in anticancer therapy. Herein, based on our previously developed DNA-mediated supercharged green fluorescent protein (ScGFP)/graphene oxide (GO) interaction, coupled with methylation-initiat...

Protein phosphorylation catalyzed by protein kinases plays a critical role in many intracellular processes, and detecting kinase activity is important in biochemical research and drug discovery. Herein, we developed a novel fluorescent biosensor to detect protein kinase activity based on phosphorylation-mediated assembly of semisynthetic green fluo...

Nowadays, large-scale screening for enzyme discovery, engineering and drug discovery process requires simple, fast and sensitive enzyme activity assay platforms with high integration and potential for high-throughput detection. Herein, a novel Automatic and Integrated Micro-Enzyme Assay (AIμEA) platform was proposed based on a unique micro-reaction...

Protein engineering by resurfacing is an efficient approach to provide new molecular toolkits for biotechnology and bio-analytical chemistry. H39GFP is a new variant of green fluorescent protein containing 39 histidine residues in the primary sequence, which was developed by protein resurfacing. Herein, taking H39GFP as the signal reporter, a label...

Herein, we report a fluorometric method for monitoring the activity and inhibition of protein kinase based on positively charged gold nanoparticles, (+)AuNPs. In this assay, when the cationic substrate peptide (S-peptide) is phosphorylated by protein kinase, the resulting negatively charged product peptide (P-peptide) will be adsorbed onto (+)AuNPs...

Protein kinase plays a vital role in regulating signal-transduction pathways and its simple and quick detection is highly desirable because traditional kinase assays typically rely on a time-consuming kinase-phosphorylation process (ca. 1 h). Herein, we report a new and rapid fluorescence-based sensing platform for probing the activity of protein k...

Supercharged proteins are a new class of functional proteins with exceptional stability and potent ability to deliver bio-macromolecules into cells. As a proof-of-principle, a novel application of supercharged proteins as a versatile biosensing platform for nucleic acid detection and epigenetics analysis is presented. Taking supercharged green fluo...

Herein, a novel label-free fluorescent assay has been developed to detect the activity of thrombin and its inhibitor, based on a recombinant enhanced green fluorescence protein (EGFP) and Ni(2+) ions immobilized nitrilotriacetic acid-coated magnetic nanoparticles (Ni(2+)-NTA MNPs). The EGFP, containing a thrombin cleavage site and a hexahistidine s...

We report here an affinity separation-based fluorometric method for monitoring the activity and inhibition of protein kinase. In this assay, when the fluorescein isothiocyanate (FITC) labeled substrate peptides (S-peptide) are phosphorylated by kinase, the product peptides (P-peptide) will be adsorbed and concentrated onto the surface of Zr(4+)-imm...

The rapid development in nanoparticle production and application during the past decade requires an easy, rapid, and predictive screening method for nanoparticles toxicity assay. In this study, the toxicological effects and the source of toxicity of copper nanoparticles (CuNPs) are investigated based on a stress-responsive bacterial biosensor array...

Cystatin C (Cys C) is a significant cysteine protease inhibitor in human bodies, and is proposed as a fascinating novel marker of glomerular filtration rate for kidney injury detection. Almost all traditional methods for Cys C measurement are immunoassays. In this article, we report a simple, immune-independent (no need to rely on immunoassay) and...

Study on antioxidants' radical scavenging processes and antioxidant capabilities is important for understanding the protective role of antioxidants against oxidative damages associated with some chronic diseases and food degradation. Traditional methods to monitor the radical scavenging by antioxidant require expensive instrument and sophisticated...

A simple and ultrasensitive label-free electrochemical impedimetric aptasensor for thrombin based on the cascaded signal amplification was reported. The sandwich system of aptamer/thrombin/aptamer-functionalized Au nanoparticles (Apt-AuNPs) was fabricated as the sensing platform. The change of the interfacial feature of the electrode was characteri...