ArticleAdvanced science (Weinheim, Baden-Wurttemberg, Germany)2021
Hairpin-Spacer crRNA-Enhanced CRISPR/Cas13a System Promotes the Specificity of Single Nucleotide Polymorphism (SNP) Identification.
Article in Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed.
- Repeat region engineering of Cas13a crRNA enables conformational gating-based autocatalytic CRISPR biosensing.Nucleic acids research · 2026Article
- Cas-regulation-targeting chimera enables selective and tunable control of CRISPR/Cas12a.Nucleic acids research · 2026Article
- Molecular engineering of CRISPR/Cas12a: from activity enhancement to exponential signal amplification.Chemical science · 2026Review
- A kinetic approach for mapping seed regions of CRISPR ribonucleoprotein and improving specificity.Nucleic acids research · 2026Article
- Advanced CRISPR Technologies for RNA Imaging in Live Cells.Chemical & biomedical imaging · 2026Review
- A novel Dual-guide CRISPR-Cas13 strategy improves specificity for single-nucleotide variant detection.Nucleic acids research · 2026Article
- Recent advances in CRISPR-based single-nucleotide fidelity diagnostics.Communications medicine · 2025Review
- Detection of Murine Norovirus on Fresh Produce Through a CRISPR/Cas13a RNase-Based Capsid Integrity Assay.Food and environmental virology · 2025Article
- Topologically constrained DNA-mediated one-pot CRISPR assay for rapid detection of viral RNA with single nucleotide resolution.EBioMedicine · 2025Article
- CRISPR detection of cardiac tumor-associated microRNAs.Molecular biology reports · 2025Review
- Harnessing noncanonical trans-cleavage characteristics of Cas12 and Cas13a to enhance CRISPR-based diagnostics.Communications biology · 2024Article
- Integrating allele-specific PCR with CRISPR-Cas13a for sensitive KRAS mutation detection in pancreatic cancer.Journal of biological engineering · 2024Article
- CRISPR-Cas target recognition for sensing viral and cancer biomarkers.Nucleic acids research · 2024Review
- Dynamic Addressing Molecular Robot (DAMR): An Effective and Efficient Trial-and-Error Approach for the Analysis of Single Nucleotide Polymorphisms.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- CRISPR/Cas13a-based supersensitive circulating tumor DNA assay for detecting EGFR mutations in plasma.Communications biology · 2024Article
- A high-fidelity DNAzyme-assisted CRISPR/Cas13a system with single-nucleotide resolved specificity.Chemical science · 2024Article
- PCR Independent Strategy-Based Biosensors for RNA Detection.Biosensors · 2024Review
- Programmable RNA targeting with CRISPR-Cas13.RNA biology · 2024Review
- Advances in application of CRISPR-Cas13a system.Frontiers in cellular and infection microbiology · 2024Review
- Unmodificated stepless regulation of CRISPR/Cas12a multi-performance.Nucleic acids research · 2023Article
Corrections and comments
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Authors and funding
14 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Cas13a system has great potential in RNA interference and molecular diagnostic fields. However, lacking guidelines for crRNA design hinders practical applications of the Cas13a system in RNA editing and single nucleotide polymorphism identification. This study posits that crRNAs with hairpin spacers improve the specificity of CRISPR/Cas13a system (termed hs-CRISPR). Gibbs free energy analysis suggests that the hairpin-spacer crRNAs (hs-crRNAs) suppress Cas13a's affinity to off-target RNA. A hepatitis B virus DNA genotyping platform is established to further validate the high-specificity of hs-CRISPR/Cas13a system. Compared to ordinary crRNA, hs-crRNAs increase the specificity by threefold without sacrificing the sensitivity of the CRISPR/Cas13a system. Furthermore, the mechanism of the Cas13a/hs-crRNA/target RNA composition is elucidated with theoretical simulations. This work builds on the fundamental understanding of Cas13a activation and offers significant improvements for the rational design of crRNA for the CRISPR/Cas13a system.
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