ArticleChemical science2022
PAM-less conditional DNA substrates leverage trans-cleavage of CRISPR-Cas12a for versatile live-cell biosensing.
Article in Chemical science, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- Recent advances in CRISPR/Cas12a-based biosensors for heavy metal analysis: a review of signal readout strategies and analytical performance.Analytical and bioanalytical chemistry · 2026Review
- RAPID: evaluation of Cas12a protospacer nicking and chimeric reporters for PAM-independent RNA and DNA diagnostics.Nucleic acids research · 2026Article
- CRISPR diagnostics: from trans-nuclease activity to cancer diagnosis.Cell & bioscience · 2026Review
- Amplification-Free Dual-Blocking Autocatalytic CRISPR-Cascade for Atto-Molar DNA Detection with Low Nonspecific Signal.bioRxiv : the preprint server for biology · 2025Article
- CRISPR-Cas-Based Diagnostics in Biomedicine: Principles, Applications, and Future Trajectories.Biosensors · 2025Review
- Spatially blocked split CRISPR-Cas12a system for ultra-sensitive and versatile small molecule activation and detection.Nature communications · 2025Article
- Exploring the effect of activator topology on CRISPR-Cas12a trans-cleavage activity.Nucleic acids research · 2025Article
- Engineering stimuli-responsive CRISPR-Cas systems for versatile biosensing.Analytical and bioanalytical chemistry · 2025Review
- PAM-Independent CRISPR-Cas12a System for Specific Assays of Single Nucleotide Variants.JACS Au · 2025Article
- Regulation of CRISPR trans-cleavage activity by an overhanging activator.Nucleic acids research · 2025Article
- Binding-driven forward tearing protospacer activated CRISPR-Cas12a system and applications for microRNA detection.Journal of nanobiotechnology · 2024Article
- Recent Advances in the CRISPR/Cas-Based Nucleic Acid Biosensor for Food Analysis: A Review.Foods (Basel, Switzerland) · 2024Review
- Light-activated CRISPR-Cas12a for amplified imaging of microRNA in cell cycle phases at single-cell levels.Science advances · 2024Article
- A high-fidelity DNAzyme-assisted CRISPR/Cas13a system with single-nucleotide resolved specificity.Chemical science · 2024Article
- 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
- In vivo imaging of mitochondrial DNA mutations using an integrated nano Cas12a sensor.Nature communications · 2023Article
- A collagen-immobilized nanodevice forChemical science · 2023Article
- Signal Amplification by theAnalytical chemistry · 2023Review
- PAM-independent ultra-specific activation of CRISPR-Cas12a via sticky-end dsDNA.Nucleic acids research · 2022Article
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The CRISPR-Cas system has been repurposed as a powerful live-cell imaging tool, but its utility is limited to genomic loci and mRNA imaging in living cells. Here, we demonstrated the potential of the CRISPR-Cas system as a generalizable live-cell biosensing tool by extending its applicability to monitor diverse intracellular biomolecules. In this work, we engineered a CRISPR-Cas12a system with a generalized stimulus-responsive switch mechanism based on PAM-less conditional DNA substrates (pcDNAs). The pcDNAs with stimulus-responsiveness toward a trigger were constructed from the DNA substrates featuring no requirement of a protospacer-adjacent motif (PAM) and a bubble structure. With further leveraging the trans-cleavage activity of CRISPR-Cas12a for signal reporting, we established a versatile CRISPR-based live-cell biosensing system. This system enabled the sensitive sensing of various intracellular biomolecules, such as telomerase, ATP, and microRNA-21, making it a helpful tool for basic biochemical research and disease diagnostics.
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Registered trials
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