Protein-Scaffolded DNA Nanostructures for Imaging of Apurinic/Apyrimidinic Endonuclease 1 Activity in Live Cells

Li, Junjie; Du, Wenwen; Liu, Yining; Wang, Fenglin; Tang, Li‐Juan; Jiang, Jian‐Hui

doi:10.1021/acs.analchem.2c05504

Cited by 9 publications

(3 citation statements)

References 26 publications

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“…Subsequently, the endogenously stimulated AAE system was investigated for cell-selective imaging of miRNA (Figure A). All AAE probes were delivered into three different cell lines with different expressions of APE1 and miR-21, including MCF-7 cells with overexpressed APE1 and miR-21, HeLa cells with moderate expression of APE1 and miR-21, and MCF-10A with slight expression of APE1 and miR-21. − After incubation, the fluorescence signals of these different cells were acquired by CLSM (Figure B) and are summarized in Figure C. A significant fluorescence was presented in MCF-7 cells, while a bright fluorescence was observed in HeLa cells.…”

Section: Resultsmentioning

confidence: 99%

Multiply Guaranteed Catalytic DNA Circuit for Cancer-Cell-Selective Imaging of miRNA and Robust Evaluation of Drug Resistance

Wang,

Shang,

Zhu

et al. 2024

Anal. Chem.

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Catalytic DNA circuits are desirable for sensitive bioimaging in living cells; yet, it remains a challenge to monitor these intricate signal communications because of the uncontrolled circuitry leakage and insufficient cell selectivity. Herein, a simple yet powerful DNA-repairing enzyme (APE1) activation strategy is introduced to achieve the site-specific exposure of a catalytic DNA circuit for realizing the selectively amplified imaging of intracellular microRNA and robust evaluation of the APE1-involved drug resistance. Specifically, the circuitry reactants are firmly blocked by the enzyme recognition/cleavage site to prevent undesirable off-site circuitry leakage. The caged DNA circuit has no targetsensing activity until its circuitry components are activated via the enzyme-mediated structural reconstitution and finally transduces the amplified fluorescence signal within the miRNA stimulation. The designed DNA circuit demonstrates an enhanced signal-to-background ratio of miRNA assay as compared with the conventional DNA circuit and enables the cancer-cellselective imaging of miRNA. In addition, it shows robust sensing performance in visualizing the APE1-mediated chemoresistance in living cells, which is anticipated to achieve in-depth clinical diagnosis and chemotherapy research.

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Section: Resultsmentioning

confidence: 99%

Multiply Guaranteed Catalytic DNA Circuit for Cancer-Cell-Selective Imaging of miRNA and Robust Evaluation of Drug Resistance

Wang,

Shang,

Zhu

et al. 2024

Anal. Chem.

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“…After the AP site was cleaved by APE1, it led to the restoration of the aptamer recognition ability of cytochrome c. This method allowed for the imaging of cytochrome c and APE1 with a DNA probe, which provided a basis for the detection of biological functions of related proteins. Inspired by the strong interaction between avidin and APE1, Li et al proposed a DNA nanostructure, which consisted of a protein (avidin) as a connecting structure, for imaging the activity of APE1 in different live cells (shown in Figure 8) [99]. The avidin-scaffolded DNA nanostructure, which was prepared based on the interaction between avidin and biotin, had a relatively loose structure with little steric hindrance.…”

Section: Dna/organic Nanomaterials Compositesmentioning

confidence: 99%

DNA Nanotechnology-Empowered Fluorescence Imaging of APE1 Activity

He,

Liu,

et al. 2023

Chemistry

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Apurinic/apyrimidinic endonuclease 1 (APE1), also known as redox factor-1 (Ref-1), is a multifunctional protein that exists widely in living organisms. It can specifically recognize and cleave the DNA in apurinic/apyrimidinic (AP) sites in the base excision repair (BER) pathway, as well as regulate the expression of genes to activate some transcription factors. The abnormal expression and disruptions in the biological functions of APE1 are linked to a number of diseases, including inflammation, immunodeficiency, and cancer. Hence, it is extremely desired to monitor the activity of APE1, acquiring a thorough understanding of the healing process of damaged DNA and making clinical diagnoses. Thanks to the advent of DNA nanotechnology, some nanodevices are used to image the activity of APE1 with great sensitivity and simplicity. In this review, we will summarize developments in DNA-nanotechnology-empowered fluorescence imaging in recent years for APE1 activity according to different types of DNA probes, which are classified into linear DNA probes, composite DNA nanomaterials, and three-dimensional (3D) DNA nanostructures. We also highlight the future research directions in the field of APE1 activity imaging.

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“…In situ imaging of disease-related biomarkers in living cells for the identification of tumor cells has the advantages of simple operation, no need for complex sample pretreatment, and can obtain more effective information, promoting accurate early diagnosis and treatment of tumors . In situ imaging disease-related biomarkers, such as ions, small molecules, proteins, and RNA, have seen significant progress and sparked widespread interest. Recently, many efforts have been made to achieve specific discrimination of cells. , However, the rapid and accurate identification of tumor cells by imaging disease-related biomarkers remains a challenge due to some non-negligible drawbacks.…”

Section: Introductionmentioning

confidence: 99%

Dual miRNA-Triggered DNA Walker Assisted by APE1 for Specific Recognition of Tumor Cells

Zhou,

Li,

et al. 2024

Anal. Chem.

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The identification of a specific tumor cell is crucial for the early diagnosis and treatment of cancer. However, it remains a challenge due to the limited sensitivity and accuracy, long response time, and low contrast of the recent approaches. In this study, we develop a dual miRNAtriggered DNA walker (DMTDW) assisted by APE1 for the specific recognition of tumor cells. miR-10b and miR-155 were selected as the research models. Without miR-10b and miR-155 presence, the DNA walker remains inactive as its walking strand of W is locked by L1 and L2. After miR-10b and miR-155 are input, the DNA walker is triggered as miR-10b and miR-155 bind to L1 and L2 of W-L1-L2, respectively, unlocking W. The DNA walker is driven by endogenous APE1 that is highly catalytic and is highly expressed in the cytoplasm of tumor cells but barely expressed in normal cells, ensuring high contrast and reaction efficiency for specific recognition of tumor cells. Dual miRNA input is required to trigger the DNA walker, making this strategy with a high accuracy. The DMTDW strategy exhibited high sensitivity for miRNA analysis with a detection limit of 44.05 pM. Living cell-imaging experiments confirmed that the DMTDW could effectively respond to the fluctuation of miRNA and specifically identified MDA-MB-231 cells from different cell lines. The proposed DMTDW is sensitive, rapid, and accurate for specific tumor cell recognition. We believe that the DMTDW strategy can become a powerful diagnostic tool for the specific recognition of tumor cells.

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Protein-Scaffolded DNA Nanostructures for Imaging of Apurinic/Apyrimidinic Endonuclease 1 Activity in Live Cells

Cited by 9 publications

References 26 publications

Multiply Guaranteed Catalytic DNA Circuit for Cancer-Cell-Selective Imaging of miRNA and Robust Evaluation of Drug Resistance

Multiply Guaranteed Catalytic DNA Circuit for Cancer-Cell-Selective Imaging of miRNA and Robust Evaluation of Drug Resistance

DNA Nanotechnology-Empowered Fluorescence Imaging of APE1 Activity

Dual miRNA-Triggered DNA Walker Assisted by APE1 for Specific Recognition of Tumor Cells

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