Construction of a dual-model aptasensor based on G-quadruplexes generated via rolling circle amplification for visual/sensitive detection of kanamycin

Gao, Xiaolin; Sun, Zhicong; Wang, Xiaoyang; Zhang, Wanqi; Xu, Deyan; Sun, Xia; Guo, Yemin; Xu, Shicai; Li, Falan

doi:10.1016/j.scitotenv.2022.156276

Cited by 19 publications

(4 citation statements)

References 33 publications

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“…Subsequently, a conventional 3,3′,5,5′-tetramethylbenzidine (TMB)-based colorimetric assay was employed to validate the sustained HRP activity within the HRP-Apt conjugate (Figure S2). 29 Having successfully prepared HRP-Apt, we proceeded to investigate the characteristics of affinity-primed proximity labeling by employing HRP-Apt. Biotin−phenol is a commonly used labeling molecule in such reactions to demonstrate the local labeling effect.…”

Section: ■ Introductionmentioning

confidence: 99%

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Highly Sensitive Labeling, Clickable Functionalization, and Glycoengineering of the MUC1 Neighboring System

Wang,

Chen,

Wei

et al. 2024

JACS Au

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This study introduces a novel wash-type affinity-primed proximity labeling (WAPL) strategy for labeling and surface engineering of the MUC1 protein neighboring system. The strategy entails the utilization of peroxidase in conjunction with a MUC1-selective aptamer, facilitating targeted binding to MUC1 and inducing covalent labeling of the protein neighboring system. This study reveals a novel finding that the WAPL strategy demonstrates superior labeling efficiency in comparison to nonwash-type affinity-primed proximity labeling, marking the first instance of such observations. The WAPL strategy provides signal amplification by converting a single recognition event into multiple covalent labeling events, thereby improving the detection sensitivity for subtle changes in MUC1. The WAPL platform employs two levels of labeling upgrades, modifying the biotin handles of the conventional labeling substrate, biotin−phenol. The first level involves a range of clickable molecules, facilitating dibenzoazacyclooctynylation, alkynylation, and trans-cyclooctenylation of the protein neighboring system. The second level utilizes lactose as a post-translational modification model, enabling rapid and reliable glycoengineering of the MUC1 neighboring system while remaining compatible with cell-based assays. The implementation of the WAPL strategy in protein neighboring systems has resulted in the establishment of a versatile platform that can effectively facilitate diverse monitoring and regulation techniques. This platform offers valuable insights into the regulation of relevant signaling pathways and promotes the advancement of novel therapeutic approaches, thereby bringing substantial implications for human health.

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Section: ■ Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Highly Sensitive Labeling, Clickable Functionalization, and Glycoengineering of the MUC1 Neighboring System

Wang,

Chen,

Wei

et al. 2024

JACS Au

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“…DNAzyme is a functional DNA oligonucleotide that resembles a biological enzyme . G-quadruplex/hemin (G4/hemin) DNAzyme mimics the catalytic activity of horseradish peroxidase by wrapping G-quadruplex structure around hemin, and can catalyze ABTS, TMB, and tyramine for colorimetric or fluorescence detection in the presence of H 2 O 2 . − With good thermal stability, low cost, and easy operation, it has promising application prospects in bioanalytical and biomedical fields. − However, the catalytic activity of traditional G4/hemin DNAzyme is relatively poor for low abundance target detection and susceptible to complex environmental influence. − Free hemin monomers that are not wrapped by G-quadruplex can also catalyze the peroxidation reaction to a certain extent, leading to background signal leakage . In our previous work, an innovative proximity-enhanced cofactor assembly strategy was proposed.…”

Section: Introductionmentioning

confidence: 99%

Visual Assay for Methicillin-Resistant Staphylococcus aureus Based on Rolling Circular Amplification Triggering G-Quadruplex/Hemin DNAzyme Proximity Assembly

et al. 2023

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Nowadays, infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have constituted a new challenge for anti-infective treatment. Precise identification and rapid clinical diagnostics of MRSA from other methicillin-sensitive strains entail assays with robust diagnostic efficiency and simple operation steps. Sensitive detection of MecA gene is promising to indicate MRSA infection, but it is challenged by the lack of isothermal and simple strategies. A visual assay based on isothermal rolling circular amplification and G-quadruplex/hemin (G4/hemin) DNAzyme proximity assembly was proposed for the immediate, efficient, and cost-effective detection of MecA in simple operation steps and in a single tube. The presence of MecA specifically drove the formation of circular templates, which further triggered isothermal amplification. The amplified product offered abundant binding sites for DNA-grafted hemin probes to form a novel proximity-assembled G4/hemin DNAzyme structure for colorimetric changing diagnosis. This tandem-repeated novel DNAzyme possessed higher catalytic activity and a lower background signal than traditional G4/hemin DNAzyme, ensuring sensitive discrimination of MRSA (limit of detection: 9.6 pM). Assay stability and antimatrix interference capability enable clinical application, which shows compared diagnostic ability with classic methods (100% sensitivity and 100% specificity) but possesses more simplified procedures and shorter turnaround time (<6 h). This colorimetric strategy in a nonsite-specific and hypersensitive manner holds foreseeable prospects in clinical diagnostic and research applications.

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“…By changing the target recognition regions, padlock probes can target different nucleic acid molecules. 8 Moreover, by rationally designing the functional region, RCA products can output signals in various ways, such as by forming G quadruplexes, combining with hemin, thioflavin, or zinc phthalocyanine to catalyze the color development of substrates, or forming photoelectric signals, 9,10 triggering hyperbranched RCA (HRCA), and outputting fluorescence signals through the intercalation of fluorescent dyes. 11 Numerous studies have achieved effective nucleic acid detection using padlock-assisted RCA because of its advantages of efficient isothermal amplification, easy editing, and flexible signal output.…”

Section: Introductionmentioning

confidence: 99%

A Universal Strategy for Enhancing the Circulating miRNAs’ Detection Performance of Rolling Circle Amplification by Using a Dual-Terminal Stem-Loop Padlock

Xu,

Wu,

Liu

et al. 2023

ACS Nano

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Rolling circle amplification (RCA) is one of the most promising nucleic acid detection technologies and has been widely used in the molecular diagnosis of disease. Padlock probes are often used to form circular templates, which are the core of RCA. However, RCA often suffers from insufficient specificity and sensitivity. Here we report a reconstruction strategy for conventional padlock probes to promote their overall performance in nucleic acid detection while maintaining probe functions uncompromised. When two rationally designed stem-loops were strategically placed at the two terminals of linear padlock probes, the specificity of target recognition was enhanced and the negative signal was significantly delayed. Our design achieved the best single-base discrimination compared with other structures and over a 1000-fold higher sensitivity than that of the conventional padlock probe, validating the effectiveness of this reconstruction. In addition, the underlying mechanisms of our design were elucidated through molecular dynamics simulations, and the versatility was validated with longer and shorter padlocks targeting the same target, as well as five additional targets (four miRNAs and dengue virus – 2 RNA mimic (DENV-2)). Finally, clinical applicability in multiplex detection was demonstrated by testing real plasma samples. Our exploration of the structures of nucleic acids provided another perspective for developing high-performance detection systems, improving the efficacy of practical detection strategies, and advancing clinical diagnostic research.

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Construction of a dual-model aptasensor based on G-quadruplexes generated via rolling circle amplification for visual/sensitive detection of kanamycin

Cited by 19 publications

References 33 publications

Highly Sensitive Labeling, Clickable Functionalization, and Glycoengineering of the MUC1 Neighboring System

Highly Sensitive Labeling, Clickable Functionalization, and Glycoengineering of the MUC1 Neighboring System

Visual Assay for Methicillin-Resistant Staphylococcus aureus Based on Rolling Circular Amplification Triggering G-Quadruplex/Hemin DNAzyme Proximity Assembly

A Universal Strategy for Enhancing the Circulating miRNAs’ Detection Performance of Rolling Circle Amplification by Using a Dual-Terminal Stem-Loop Padlock

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