Label-free chemiluminescent CRISPR/Cas12a biosensing strategy for the detection of nucleic acids and non-nucleic acids utilizing DNAzyme Supernova

Qin, Cheng; Bian, Xinlan; Lv, Wei; Li, Bingzhi

doi:10.1016/j.snb.2023.135143

Cited by 5 publications

(2 citation statements)

References 43 publications

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“…162 Using this DNAzyme, Qin et al created a biosensor to identify synthetic DNA associated with the SARS-CoV-2 N gene and the monkeypox virus (MPXV) F3L gene with LODs of 1.5 and 2.4 pM, respectively. 161 They did this by integrating the Supernova DNAzyme with CRISPR/Cas12a, which can be activated in the presence of a target to rapidly cleave the ssDNA domain of Supernova, leading to an attenuation of the blue light signal.…”

Section: Other Dnazymementioning

confidence: 99%

Recent progress on DNAzyme-based biosensors for pathogen detection

Liu,

Yuan,

Xiao

2024

Anal. Methods

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Section: Other Dnazymementioning

confidence: 99%

Recent progress on DNAzyme-based biosensors for pathogen detection

Liu,

Yuan,

Xiao

2024

Anal. Methods

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“…Commonly employed Cas proteins in diagnosis encompass Cas9, Cas12a, Cas13, and Cas14. − Cas12a, among these, is activated by DNA and exhibits potential applications in diagnostics . The realm of CRISPR/Cas12a-based biosensing has garnered substantial interest, spanning from nucleic acid to non-nucleic acid targets including ions, small molecules, and even cells, to name but a few. − This technology offers advantages, including simplicity, environmental resilience, high sensitivity, and specificity. While it has rapidly progressed, its broader clinical implementation demands enhanced cost-effectiveness and accessibility.…”

Section: Introductionmentioning

confidence: 99%

Postamplifying Cas12a Activation through Hybridization Chain Reaction-Triggered Fluorescent Nanocluster Formation for Ultrasensitive Nucleic Acid Detection

Yang,

Xie,

Zhou

et al. 2024

ACS Appl. Mater. Interfaces

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CRISPR/Cas12a-based biosensing is advancing rapidly; however, achieving sensitive and cost-effective reporting of Cas12a activation remains a challenge. In response, we have developed a label-free system capable of postamplifying Cas12a activation by integrating hybridization chain reaction (HCR) and DNA-copper nanoclusters (DNA-CuNCs). The trans-cleavage of Cas12a triggers a silenced HCR, leading to the in situ assembly of fluorescent DNA-CuNCs, allowing for the turn-on reporting of Cas12a activation. Without preamplification, this assay can detect DNA with a detection limit of 5 fM. Furthermore, when coupled with preamplification, the system achieves exceptional sensitivity, detecting the monkeypox virus (MPXV) plasmid at 1 copy in human serum. In a MPXV pseudovirus-based validation test, the obtained results are in agreement with those obtained by qPCR, reinforcing the robustness of this method. Our study represents the first effort to manipulate DNA-CuNC formation on HCR for highly sensitive and cost-effective reporting of Cas12a, resulting in an efficient synthetic biology-enabled sensing platform for biosafety applications.

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Simultaneous Detection of Multiple Biomarkers by Peptide Nucleic Acids‐Based Triplex Molecular Beacon in a Nanopore

Guo,

Zhang,

Ren

et al. 2024

Analysis & Sensing

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In this study, we propose an enhanced nanopore sensing strategy that utilizes a peptide nucleic acid (PNA)‐based triplex molecular beacon sensor to achieve the simultaneous detection of multiple biomarkers with a high degree of sensitivity. The sensor is a triplex switch composed of a triplex‐forming DNA strand and an oligo‐arginine‐tagged PNA strand, serving as the target recognition moiety and signal output element, respectively. Upon target binding to the recognition element of the sensor, the PNA signal output strand is released and a hybrid complex of the target‐DNA recognition strand is formed simultaneously. Due to the positive charges carried by the PNA‐Arg strands, they could be driven through the nanopore under positive electric field, effectively eliminating interferences from co‐existing target‐DNA complexes. This approach enables label‐free, one‐step detection of targets without requiring complex treatments and procedures. Leveraging the modular properties of DNA recognition strand, this method can be applied universally, and here, we successfully demonstrate its application using three SARS‐CoV‐2 related biomarkers.

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Label-free chemiluminescent CRISPR/Cas12a biosensing strategy for the detection of nucleic acids and non-nucleic acids utilizing DNAzyme Supernova

Cited by 5 publications

References 43 publications

Recent progress on DNAzyme-based biosensors for pathogen detection

Recent progress on DNAzyme-based biosensors for pathogen detection

Postamplifying Cas12a Activation through Hybridization Chain Reaction-Triggered Fluorescent Nanocluster Formation for Ultrasensitive Nucleic Acid Detection

Simultaneous Detection of Multiple Biomarkers by Peptide Nucleic Acids‐Based Triplex Molecular Beacon in a Nanopore

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