Structural insights into the AFB1 aptamer coupled with a rationally designed CRISPR/Cas12a-Exo III aptasensor for AFB1 detection

Ma, Peisheng; Guo, Hualin; Ye, Hua; Zhang, Yin; Wang, Zhouping

doi:10.1016/j.ijbiomac.2022.11.177

Cited by 24 publications

(4 citation statements)

References 38 publications

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“…As shown in Figure 4D, only AFB 1 can cause a significant change in the ΔCL value, while other analogues elicited a negligible change. The excellent selectivity of this sensor for AFB 1 detection is mainly attributed to the aptamer with a high affinity for AFB 1 and comprises a dissociation constant of less than 2.0 × 10 −6 M. 32,35 3.4. Accuracy and Reproducibility of the CRISPR/ Cas12a-Assisted Chemiluminescence Sensor.…”

Section: Resultsmentioning

confidence: 99%

“…In the case of this approach, it has several unparalleled advantages for AFB 1 detection: (1) Triple signal amplification mediated by the Cas enzyme, RCA, and HRP endows the method with high sensitivity. (2) Previous CRISPR/Cas12a-mediated aptasensors mainly include a fluorophore quencher-based reporter or nanoparticle/tag molecule-based reporter , that either suffer from relatively low sensitivity and photobleaching or were limited by the stability of the functional material. In comparison to these reported approaches, our enzyme reporter-based strategy has a relatively high sensitivity with simple signal amplification and excellent stability.…”

Section: Resultsmentioning

confidence: 99%

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CRISPR/Cas12a-Assisted Chemiluminescence Sensor for Aflatoxin B₁ Detection in Cereal Based on Functional Nucleic Acid and In-Pipet Rolling Circle Amplification

Wang

Wei

Ruan

et al. 2023

J. Agric. Food Chem.

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Herein, we report a CRISPR/Cas12a-assisted chemiluminescence sensor for aflatoxin B 1 (AFB 1 ) detection based on functional nucleic-acid-mediated target recognition and in-pipet rolling circle amplification-mediated signal amplification. In this sensor, we performed rolling circle amplification on the inside of the pipet to enrich horseradish peroxidase (pipet-poly-HRP). When AFB 1 is present, it interacts with functional nucleic acids and results in the release of the activator. The activator is designed to activate the CRISPR/Cas12a system, which cleaves the pipet-poly-HRP to liberate HRP. The freed HRP can then be measured by chemiluminescence to quantify AFB 1 . This CRISPR/Cas12a-assisted chemiluminescence sensor enables facile, highly sensitive, and specific detection of AFB 1 , with a linear range from 50 pg/mL to 100 ng/mL and a detection limit of 5.2 pg/mL. Furthermore, it exhibits satisfactory recovery and has successfully challenged AFB 1 detection in cereal samples. The proposed sensor offers a novel rapid screening approach that holds great promise for food security monitoring.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

CRISPR/Cas12a-Assisted Chemiluminescence Sensor for Aflatoxin B₁ Detection in Cereal Based on Functional Nucleic Acid and In-Pipet Rolling Circle Amplification

Wang

Wei

Ruan

et al. 2023

J. Agric. Food Chem.

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“…After optimization, the LoD for AFB1 was 0.00257 fg/mL. 60 Exo III 61 and MXenes 62 were introduced to construct a CRISPR/Cas12apowered aptasensor to detect AFB1 for the improvement of sensitivity. In the CRISPR/Cas12a-Exo III aptasensor, the Exo III-assisted cycle amplification was the first step for the enhancement of the target due to the digestion of Exo III from the 3′-end in the presence of AFB1, resulting in the release of the complementary DNA (cDNA).…”

Section: Detection Of Heavy Metal Ionsmentioning

confidence: 99%

CRISPR/Cas System: The Accelerator for the Development of Non-nucleic Acid Target Detection in Food Safety

Li,

Zhao,

Liu

et al. 2023

J. Agric. Food Chem.

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Non-nucleic acid targets have posed a serious challenge to food safety. The detection of non-nucleic acid targets can enable us to monitor food contamination in a timely manner. In recent years, the CRISPR/Cas system has been extensively explored in biosensing. However, there is a lack of a summary of CRISPR/Cas-powered detection tailored to non-nucleic acid targets involved in food safety. This review comprehensively summarizes the recent advances on the construction of CRISPR/Cas-powered detection and the promising applications in the field of food safety related non-nucleic acid targets. The current challenges and futuristic perspectives are also proposed accordingly. The rapidly evolving CRISPR/Cas system has provided a powerful propellant for nonnucleic acid target detection via integration with aptamer and/or DNAzyme. Compared with traditional analytical methods, CRISPR/Cas-powered detection is conceptually novel, essentially eliminates the dependence on large instruments, and also demonstrates the capability for rapid, accurate, sensitive, and on-site testing.

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“…[15] Mao et al have reported on upconversion-mediated CRISPR-Cas12a biosensing for the sensitive detection of OTA [16] and Ma et al have designed a CRISPR/Cas12a-Exo III aptasensor for the sensitive detection of AFB 1 . [17] Despite their excellent performance, existing CRISPR-Cas biosensing systems still present some inherent limitations, including the restriction of the protospacer adjacent motif (PAM) or protospacer flanking site (PFS) adjacent to the target sequence and the costliness and degradability of RNA. [18] In addition, the critical drawback of CRISPR-Cas is the inability to achieve one-pot, single-enzyme multiplexed assays due to its trans-cleavage activity.…”

Section: Introductionmentioning

confidence: 99%

Mesophilic Argonaute‐Based Single Polystyrene Sphere Aptamer Fluorescence Platform for the Multiplexed and Ultrasensitive Detection of Non‐Nucleic Acid Targets

Lu,

Wen,

Wang

et al. 2023

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The rapid, simultaneous, and accurate identification of multiple non‐nucleic acid targets in clinical or food samples at room temperature is essential for public health. Argonautes (Agos) are guided, programmable, target‐activated, next‐generation nucleic acid endonucleases that could realize one‐pot and multiplexed detection using a single enzyme, which cannot be achieved with CRISPR/Cas. However, currently reported thermophilic Ago‐based multi‐detection sensors are mainly employed in the detection of nucleic acids. Herein, this work proposes a Mesophilic Argonaute Report‐based single millimeter Polystyrene Sphere (MARPS) multiplex detection platform for the simultaneous analysis of non‐nucleic acid targets. The aptamer is utilized as the recognition element, and a single millimeter‐sized polystyrene sphere (PSmm) with a large concentration of guide DNA on the surface served as the microreactor. These are combined with precise Clostridium butyricum Ago (CbAgo) cleavage and exonuclease I (Exo I) signal amplification to achieve the efficient and sensitive recognition of non‐nucleic acid targets, such as mycotoxins (<60 pg mL−1) and pathogenic bacteria (<102 cfu mL−1). The novel MARPS platform is the first to use mesophilic Agos for the multiplex detection of non‐nucleic acid targets, overcoming the limitations of CRISPR/Cas in this regard and representing a major advancement in non‐nucleic acid target detection using a gene‐editing‐based system.

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Structural insights into the AFB1 aptamer coupled with a rationally designed CRISPR/Cas12a-Exo III aptasensor for AFB1 detection

Cited by 24 publications

References 38 publications

CRISPR/Cas12a-Assisted Chemiluminescence Sensor for Aflatoxin B₁ Detection in Cereal Based on Functional Nucleic Acid and In-Pipet Rolling Circle Amplification

CRISPR/Cas12a-Assisted Chemiluminescence Sensor for Aflatoxin B₁ Detection in Cereal Based on Functional Nucleic Acid and In-Pipet Rolling Circle Amplification

CRISPR/Cas System: The Accelerator for the Development of Non-nucleic Acid Target Detection in Food Safety

Mesophilic Argonaute‐Based Single Polystyrene Sphere Aptamer Fluorescence Platform for the Multiplexed and Ultrasensitive Detection of Non‐Nucleic Acid Targets

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Structural insights into the AFB1 aptamer coupled with a rationally designed CRISPR/Cas12a-Exo III aptasensor for AFB1 detection

Cited by 24 publications

References 38 publications

CRISPR/Cas12a-Assisted Chemiluminescence Sensor for Aflatoxin B1 Detection in Cereal Based on Functional Nucleic Acid and In-Pipet Rolling Circle Amplification

CRISPR/Cas12a-Assisted Chemiluminescence Sensor for Aflatoxin B1 Detection in Cereal Based on Functional Nucleic Acid and In-Pipet Rolling Circle Amplification

CRISPR/Cas System: The Accelerator for the Development of Non-nucleic Acid Target Detection in Food Safety

Mesophilic Argonaute‐Based Single Polystyrene Sphere Aptamer Fluorescence Platform for the Multiplexed and Ultrasensitive Detection of Non‐Nucleic Acid Targets

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Product

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CRISPR/Cas12a-Assisted Chemiluminescence Sensor for Aflatoxin B₁ Detection in Cereal Based on Functional Nucleic Acid and In-Pipet Rolling Circle Amplification

CRISPR/Cas12a-Assisted Chemiluminescence Sensor for Aflatoxin B₁ Detection in Cereal Based on Functional Nucleic Acid and In-Pipet Rolling Circle Amplification