CRISPR/Cas12a-based fluorescence assay for the detection of acetylcholinesterase activity

Wang, Hui-Yi; Liu, Pengfei; Hang, Xiao-Min; Zhao, Kai-Ren; Wang, Li

doi:10.1016/j.snb.2022.132691

Cited by 10 publications

(6 citation statements)

References 46 publications

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“…The amounts of Cas12a protein, crRNA, and FQ reporter were fixed according to our previous research. 41 A 23-nt ssDNA sequence we reported previously 42 was used as the initiator, and its amount was first optimized in the range of 100 nM to 5 μM (Fig. S3A †).…”

Section: Optimization Of Experimental Conditionsmentioning

confidence: 99%

A MnO₂ nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water

Yu,

Liang,

Wang

et al. 2024

Analyst

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Section: Optimization Of Experimental Conditionsmentioning

confidence: 99%

A MnO₂ nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water

Yu,

Liang,

Wang

et al. 2024

Analyst

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“…Besides, the optimum DNA probe concentration and acrylamide concentration was also by the thermogram by combining two single factors. For the different reaction time for exosomes detection (30,40,50,60, and 70 min) and temperature (21、29、37、45、53) were carried out to gain the optimal results. In addition, since the embedding amount of Cas12a/crRNA in DNA hydrogel can significantly affect the sensitivity of the analysis, the concentration of Cas12 (25, 50, 100, 150, and 200 nM) was optimized as well.…”

Section: Optimization Of the Detectionmentioning

confidence: 99%

“…In principle, these methods rely mainly on introducing the Cas12a and reporter probes after the upstream recognition. 30,31 The tedious procedures only allow the signal substance to be released first, then start their second function as a trigger in the downstream response. 32 Therefore, developing an intelligent program with "recognition-response" guided by Cas12a will simplify the analysis protocol and has potential application value to be a versatile platform for other targets.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas12a-Loaded intelligent DNA hydrogel for Universal and Ultrasensitive Exosome Assay

Luo,

Wang,

Tang

et al. 2023

Preprint

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Tumor-derived exosomes are crucial for early non-invasive and accurate tumor diagnosis in clinical. The development of highly sensitive, simple and intuitive exosome assays has sparked a research upsurge in clinical diagnostics. Here, we develop a bio-responsive the intelligent DNA hydrogel loaded with CRISPR/Cas12a for universal and ultrasensitive detection of the exosomes. The aptamer serves as the target response unit and switch of the intelligent DNA hydrogel network. The target competitively disintegrates the region of the DNA linkers then Cas12a/crRNA encapsulated in the intelligent DNA hydrogel can be released and activated by the collateral sequence, resulting in a high fluorescent intensity for exosome detection at the detection limit of 119 particles/μL. Moreover, a prefabricated colorimetric tube is made by loading a colorimetric filter membrane on the tube lid and intelligent DNA hydrogel on the tube bottom, which enables one-pot portable colorimetric detection. Without the need for laboratory instruments and professionals, this strategy allows for naked eye detection with LOD as low as 104 particles/μL, and shows great applicability in distinguishing between healthy individuals, pretreatment patients, and post treatment patients after obtaining a testable analyte. Collectively, this study constructs an ultra-sensitive detection platform for exosomes, which enables one-step sensing and dual signal output, making it a promising tool for the application of liquid biopsy based on exosomes in clinical diagnosis.

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“…Presently, the detection strategies of various biomarkers (such as N‐gene, 27 transcription factor, 28 nucleic acids 29 ) have been identified using intelligent hydrogel sensing platforms and Cas12a. In principle, these methods rely on introducing the Cas12a and reporter probes after the upstream recognition 30,31 . However, this is a two‐step procedure, as the signal substance must be released first, before triggering the downstream response 32 .…”

Section: Introductionmentioning

confidence: 99%

“…In principle, these methods rely on introducing the Cas12a and reporter probes after the upstream recognition. 30,31 However, this is a two-step procedure, as the signal substance must be released first, before triggering the downstream response. 32 Therefore, developing an intelligent program with "recognition-response" guided by Cas12a will simplify the analysis protocol and has potential clinical application value.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas12a‐loaded intelligent DNA hydrogel for universal and ultrasensitive exosome assay

Luo,

Wang,

Tang

et al. 2024

VIEW

View full text Add to dashboard Cite

Tumor‐derived exosomes are crucial for early non‐invasive and accurate tumor diagnosis in clinical diagnostics. The development of highly sensitive, simple, and intuitive exosome assays has sparked a research upsurge in clinical diagnostics. Here, we develop a bio‐responsive intelligent DNA hydrogel loaded with CRISPR/Cas12a for universal and ultrasensitive detection of the exosomes. The aptamer serves as the target response unit and switch, competitively disintegrating the region of the DNA linkers and then Cas12a/crRNA was released and activated, resulting in a high fluorescent intensity for exosome detection at the detection limit of 119 particles/μL. Moreover, a constructed colorimetric tube is made by loading a colorimetric filter membrane on the tube lid and intelligent DNA hydrogel on the tube bottom, which enables one‐pot portable colorimetric detection. Without the need for laboratory instruments and professionals, this strategy allows for naked eye detection with limit of detection as low as 104 particles/μL, and shows great applicability in distinguishing between healthy individuals, pretreatment patients, and post‐treatment patients after obtaining a testable analyte. In this study, an ultrasensitive detection platform for exosomes that enables one‐step sensing and dual signal output was introduced. The findings here suggest that this platform is a promising tool for the application of liquid biopsy based on exosomes in clinical diagnosis.

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CRISPR/Cas12a-based fluorescence assay for the detection of acetylcholinesterase activity

Cited by 10 publications

References 46 publications

A MnO₂ nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water

A MnO₂ nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water

CRISPR/Cas12a-Loaded intelligent DNA hydrogel for Universal and Ultrasensitive Exosome Assay

CRISPR/Cas12a‐loaded intelligent DNA hydrogel for universal and ultrasensitive exosome assay

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CRISPR/Cas12a-based fluorescence assay for the detection of acetylcholinesterase activity

Cited by 10 publications

References 46 publications

A MnO2 nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water

A MnO2 nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water

CRISPR/Cas12a-Loaded intelligent DNA hydrogel for Universal and Ultrasensitive Exosome Assay

CRISPR/Cas12a‐loaded intelligent DNA hydrogel for universal and ultrasensitive exosome assay

Contact Info

Product

Resources

About

A MnO₂ nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water

A MnO₂ nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water