ArticleAnalytical chemistry2022
Development of Cas12a-Based Cell-Free Small-Molecule Biosensors via Allosteric Regulation of CRISPR Array Expression.
Article in Analytical chemistry, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Translating biosynthetic gene cluster architecture into biosensor design for microbial natural product discovery.Essays in biochemistry · 2026Review
- Engineering Allosteric Transcription Factor-Based Biosensors: Advances and Prospects for Modern Food Contaminant Monitoring.Foods (Basel, Switzerland) · 2026Review
- Horseradish peroxidase as an electrochemical reporter protein for cell-free biosensors.Synthetic biology (Oxford, England) · 2026Article
- Advancements and Applications of Split Technology in CRISPR/Cas12a: Transforming Molecular Diagnostics and Biosensing.Biosensors · 2025Review
- Advancements and Prospects in DNA-Based Bioanalytical Technology for Environmental Toxicant Detection.JACS Au · 2025Review
- Engineering stimuli-responsive CRISPR-Cas systems for versatile biosensing.Analytical and bioanalytical chemistry · 2025Review
- Mitigating Antibiotic Resistance: The Utilization of CRISPR Technology in Detection.Biosensors · 2024Review
- CRISPR/Cas system and its application in the diagnosis of animal infectious diseases.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2024Review
- Modern Plant Breeding Techniques in Crop Improvement and Genetic Diversity: From Molecular Markers and Gene Editing to Artificial Intelligence-A Critical Review.Plants (Basel, Switzerland) · 2024Review
- CRISPR-HOLMES-based NADFrontiers in bioengineering and biotechnology · 2024Article
- Recent advances and challenges of the use of the CRISPR/Cas system as a non-nucleic acid molecular diagnostic.Heliyon · 2023Review
- CRISPR Assays for Disease Diagnosis: Progress to and Barriers Remaining for Clinical Applications.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Review
- Article
- Review
- Rapid and Finely-Tuned Expression for Deployable Sensing Applications.Advances in biochemical engineering/biotechnology · 2023Article
- Bio-SCAN V2: A CRISPR/dCas9-based lateral flow assay for rapid detection of theophylline.Frontiers in bioengineering and biotechnology · 2023Article
- Characterization of a thermostable Cas13 enzyme for one-pot detection of SARS-CoV-2.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cell-free biosensors can detect various molecules, thus promising to transform the landscape of diagnostics. Here, we developed a simple, rapid, sensitive, and field-deployable small-molecule detection platform based on allosteric transcription factor (aTF)-regulated expression of a clustered regularly interspaced short palindromic repeats (CRISPR) array coupled to Cas12a activity. To this end, we engineered an expression cassette harboring a T7 promoter, an aTF binding sequence, a Cas12a CRISPR array, and protospacer adjacent motif-flanked Cas12a target sequences. In the presence of the ligand, dissociation of the aTF allows transcription of the CRISPR array; this leads to activation of Cas12a collateral activity, which cleaves a single-stranded DNA linker to free a quenched fluorophore, resulting in a rapid, significant increase of fluorescence. As a proof of concept, we used TetR as the aTF to detect different tetracycline antibiotics with high sensitivity and specificity and a simple, hand-held visualizer to develop a fluorescence-based visual readout. We also adapted a mobile phone application to further simplify the interpretation of the results. Finally, we showed that the reagents could be lyophilized to facilitate storage and distribution. This detection platform represents a valuable addition to the toolbox of cell-free, CRISPR-based biosensors, with great potential for in-field deployment to detect non-nucleic acid small molecules.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.