ArticleNucleic acids research2024
New design strategies for ultra-specific CRISPR-Cas13a-based RNA detection with single-nucleotide mismatch sensitivity.
Article in Nucleic acids research, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
34 citing papers in PubMed.
- Hybridization-Based Electrochemical Biosensors for RNA Detection: Key Concepts and Insights.Sensors and actuators. B, Chemical · 2026Article
- CRISPR-Scan: Cas13a Mismatch Tolerance Enabled Ultra-Multiplexed Detection of Same-Site Indels in Acute Myeloid Leukemia.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- RNA structure modulates Cas13 activity and enables mismatch detection.Nature biotechnology · 2026Article
- Repeat region engineering of Cas13a crRNA enables conformational gating-based autocatalytic CRISPR biosensing.Nucleic acids research · 2026Article
- Structural basis for target discrimination and activation by Cas13d.Science advances · 2026Article
- A structural accessibility principle for LbuCas13a activation by noncontiguous DNA.Nucleic acids research · 2026Article
- A Dual-Port, Smartphone-Linked, Pocket-Size Fluorimeter for Rapid Molecular Diagnostic Assays at Point of Care.IEEE sensors journal · 2026Article
- Intersegmental transfers drive target search in an RNA-targeting CRISPR system.bioRxiv : the preprint server for biology · 2026Article
- CRISPR diagnostics: from trans-nuclease activity to cancer diagnosis.Cell & bioscience · 2026Review
- A universal and orthogonal safety valve for CRISPR/Cas12a without chemical modification or external stimulation.Nucleic acids research · 2026Article
- AlphaFold3-guided tracrRNA redesign yields small monomeric Cas12f RNPs.Nucleic acids research · 2026Article
- 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
- Computation and deep-learning-driven advances in CRISPR genome editing.Nature structural & molecular biology · 2026Review
- A CRISPR-based diagnostic tool to survey drug resistance in human African trypanosomiasis.Antimicrobial agents and chemotherapy · 2025Article
- Systematic evaluation of CrRNA design parameters for optimized Cas13d-mediated RNA targeting in chicken cells.Functional & integrative genomics · 2025Article
- CRISPR/Cas12a2 enables ultra-sensitive amplification-free RNA detection.Nucleic acids research · 2025Article
- Mechanistic insights into Cas13d enzymes from cryo-EM structures of CasRx and DjCas13d.Nucleic acids research · 2025Article
- Recent advances in CRISPR-based single-nucleotide fidelity diagnostics.Communications medicine · 2025Review
Corrections and comments
- Update of
Authors and funding
10 authors.
Funding
Abstract
An increasingly pressing need for clinical diagnostics has required the development of novel nucleic acid-based detection technologies that are sensitive, fast, and inexpensive, and that can be deployed at point-of-care. Recently, the RNA-guided ribonuclease CRISPR-Cas13 has been successfully harnessed for such purposes. However, developing assays for detection of genetic variability, for example single-nucleotide polymorphisms, is still challenging and previously described design strategies are not always generalizable. Here, we expanded our characterization of LbuCas13a RNA-detection specificity by performing a combination of experimental RNA mismatch tolerance profiling, molecular dynamics simulations, protein, and crRNA engineering. We found certain positions in the crRNA-target-RNA duplex that are particularly sensitive to mismatches and establish the effect of RNA concentration in mismatch tolerance. Additionally, we determined that shortening the crRNA spacer or modifying the direct repeat of the crRNA leads to stricter specificities. Furthermore, we harnessed our understanding of LbuCas13a allosteric activation pathways through molecular dynamics and structure-guided engineering to develop novel Cas13a variants that display increased sensitivities to single-nucleotide mismatches. We deployed these Cas13a variants and crRNA design strategies to achieve superior discrimination of SARS-CoV-2 strains compared to wild-type LbuCas13a. Together, our work provides new design criteria and Cas13a variants to use in future easier-to-implement Cas13-based RNA detection applications.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
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.