A dual signal amplification method for miR-204 assay by combining chimeric molecular beacon with double-stranded nuclease

Liu, Hao; Fan, Jialong; Liu, Weiping; Tong, Chunyi; Xie, Zhenhua; Deng, Rilin; Long, Xiaoyan

doi:10.1039/c8ay02147c

Cited by 12 publications

(6 citation statements)

References 30 publications

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“…[36] DSN can effectively cleave dsDNA or hybrid double strands of DNA and RNA. [37] Xia et al constructed an aggregation-induced emission luminogens (AIEgens) functional nucleic acid probe (AFNAs) without labelled quencher group to detect intracellular Hg 2 + through DSN-assisted linear amplification with high sensitivity. [23] Using the advantage of DNA potent binding affinities with Hg 2 + , AFNAs changed the conformation from single-stranded DNA (ssDNA) to double-stranded DNA (dsDNA), producing weak fluorescence.…”

Section: Nuclease-mediated Cyclic Signal Amplificationmentioning

confidence: 99%

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Intracellular Signal Amplification for Ultrasensitive Detection and Imaging: Progress, Challenges, and Opportunities

Gong

Shan

et al. 2022

Analysis & Sensing

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Disease signaling molecules and biomarkers are important in biosensing and biomedicine, and their ultra‐sensitive detection and imaging are critical in medical diagnosis. However, most biomarkers have low levels of expression in cells and are easily degraded; therefore, they cannot be accurately detected in real time by conventional amplification methods, which are typically confined to cell lysates. To achieve highly sensitive detection of intracellular low‐abundance molecules, extensive efforts have been devoted to the development of in situ signal amplification techniques. This review tracks the development of recent advances in in situ signal amplification strategies, and systematically summarizes the advantages and disadvantages of enzymatic and non‐enzymatic amplification techniques for detecting and tracing intracellular targets. Further, perspectives, challenges, and opportunities for intracellular signal amplification are discussed to promote its adoption for in situ signal amplification in clinical settings and to provide new insights into signaling molecules and biomarkers in cellular functions and associated diseases.

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Section: Nuclease-mediated Cyclic Signal Amplificationmentioning

confidence: 99%

“…To address this issue, DSN has been gradually applied for signal amplification detection [36] . DSN can effectively cleave dsDNA or hybrid double strands of DNA and RNA [37] . Xia et al .…”

Section: Enzymatic Amplificationmentioning

confidence: 99%

Intracellular Signal Amplification for Ultrasensitive Detection and Imaging: Progress, Challenges, and Opportunities

Gong

Shan

et al. 2022

Analysis & Sensing

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“…DSN was used to increase the sensitivity for detecting miRNAs and reduce the amount of material consumed by increasing the target to probe hybridization numbers [ 70 ]. Liu et al [ 71 ] combined the target specificity of chimeric MB with DSN signal amplification capability for glioma biomarker miRNA-204 detection. They designed MB with an RNA stem that comprised a loop complementary to the target sequence.…”

Section: Diagnostic and Prognostic Of Mirna Biomarkersmentioning

confidence: 99%

State-of-the-Art Fluorescent Probes: Duplex-Specific Nuclease-Based Strategies for Early Disease Diagnostics

et al. 2022

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Precision healthcare aims to improve patient health by integrating prevention measures with early disease detection for prompt treatments. For the delivery of preventive healthcare, cutting-edge diagnostics that enable early disease detection must be clinically adopted. Duplex-specific nuclease (DSN) is a useful tool for bioanalysis since it can precisely digest DNA contained in duplexes. DSN is commonly used in biomedical and life science applications, including the construction of cDNA libraries, detection of microRNA, and single-nucleotide polymorphism (SNP) recognition. Herein, following the comprehensive introduction to the field, we highlight the clinical applicability, multi-analyte miRNA, and SNP clinical assays for disease diagnosis through large-cohort studies using DSN-based fluorescent methods. In fluorescent platforms, the signal is produced based on the probe (dyes, TaqMan, or molecular beacon) properties in proportion to the target concentration. We outline the reported fluorescent biosensors for SNP detection in the next section. This review aims to capture current knowledge of the overlapping miRNAs and SNPs’ detection that have been widely associated with the pathophysiology of cancer, cardiovascular, neural, and viral diseases. We further highlight the proficiency of DSN-based approaches in complex biological matrices or those constructed on novel nano-architectures. The outlooks on the progress in this field are discussed.

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“…The molecular beacon (MB), first proposed by Tyagi and Kramer in 1996, [24] are stem‐loop oligonucleotides labeled with a fluorophore and a quencher at the 5’ or 3’ ends [25] . MBs have become a kind of DNA probes that is widely used in various analysis targets such as thrombin, [26] transcription factors, [27] single nucleotide polymorphisms, [28,29] message‐RNA (mRNA), [30] heavy metal ions, [31,32] microRNA(miRNA) and others [33–37] . A “turn‐off” fluorescent method for sensing triplex DNA binders was reported [38] .…”

Section: Introductionmentioning

confidence: 99%

A Novel “Turn‐On” Fluorescent Sensor for Screening Triplex DNA Binder Based upon Molecular Beacon

Chen

et al. 2022

ChemistrySelect

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Triplex DNA plays an essential role in molecular recognition, gene regulation, diagnosis, and treatment of genetic diseases. However, triplex DNA has some shortcomings, such as poor internal Hoogsteen hydrogen bond stability and the difficult formation of parallel triplex DNA under physiological conditions, limiting its applications to a certain extent. Therefore, it is crucial to screen intercalators to enhance triplex DNA stability. Herein, we established a novel "turn-on" fluorescence sensor for screening triplex DNA binders based upon molecular beacon. Eight different organic small molecules were screened. The fluorescence of the sensor was significantly enhanced in the presence of berberine, coralyne, palmatine, and neomycin. These four small molecules affect stabilizing triplex DNA as an intercalator; the other four small molecules, kanamycin, tetracycline, ethidium bromide, and 4 ', 6-diamidino-2-phenylindole did not show any noticeable intercalation effect. The results of circular dichroism and melting curve were consistent with those of the fluorescence spectrum.

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A dual signal amplification method for miR-204 assay by combining chimeric molecular beacon with double-stranded nuclease

Abstract: Due to the miRNA's key roles in early disease diagnosis, an assay based on chimeric molecular beacon was constructed for miR204 detection with LOD lower to 50 pM.

Cited by 12 publications

References 30 publications

Intracellular Signal Amplification for Ultrasensitive Detection and Imaging: Progress, Challenges, and Opportunities

Intracellular Signal Amplification for Ultrasensitive Detection and Imaging: Progress, Challenges, and Opportunities

State-of-the-Art Fluorescent Probes: Duplex-Specific Nuclease-Based Strategies for Early Disease Diagnostics

A Novel “Turn‐On” Fluorescent Sensor for Screening Triplex DNA Binder Based upon Molecular Beacon

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