Trend AnalysisBiology & Life Sciences

CRISPR Diagnostics: From Gene Editing Tool to Point-of-Care Pathogen Detector

CRISPR-Cas systems were developed for genome editing, but a serendipitous property β€” the "collateral cleavage" activity of Cas12 and Cas13, where target recognition triggers non-specific degradation o...

By Sean K.S. Shin
This blog summarizes research trends based on published paper abstracts. Specific numbers or findings may contain inaccuracies. For scholarly rigor, always consult the original papers cited in each post.

The Question

CRISPR-Cas systems were developed for genome editing, but a serendipitous property β€” the "collateral cleavage" activity of Cas12 and Cas13, where target recognition triggers non-specific degradation of nearby nucleic acids β€” has been repurposed for diagnostics. SHERLOCK (Cas13-based) and DETECTR (Cas12-based) can detect specific DNA or RNA sequences with attomolar sensitivity and single-nucleotide specificity, then report detection via fluorescence or lateral flow strips visible to the naked eye. Can CRISPR diagnostics deliver PCR-level accuracy in a format deployable at point-of-care without laboratory infrastructure?

Landscape

K. Zhang et al. (2024) developed a dual lateral flow assay that simultaneously detects HPV16 and HPV18 using RPA (recombinase polymerase amplification) coupled with CRISPR-Cas. The two-pathogen, single-test format addresses a practical need: cervical cancer screening requires testing for both high-risk HPV types. Their assay achieves results in ~40 minutes with visual readout β€” eliminating the need for thermal cyclers, fluorimeters, or trained operators.

D. You et al. (2024) demonstrated a one-pot system combining isothermal amplification and CRISPR/Cas13d detection in a single tube β€” eliminating the contamination risk of transferring amplified product between tubes. One-pot operation is critical for field deployment where operator error must be minimised.

J. Chen et al. (2024) reviewed the rapidly expanding landscape of CRISPR-based visual detection methods, cataloguing innovations in signal amplification (catalytic hairpin assembly, rolling circle amplification), readout modalities (fluorescence, colorimetric, electrochemical, lateral flow), and multiplexing strategies (simultaneous detection of multiple targets).

Key Claims & Evidence

<
ClaimEvidenceVerdict
CRISPR diagnostics achieve PCR-level sensitivityAttomolar detection documented across platforms (J. Chen et al. 2024)Supported; sensitivity depends on preamplification step
Lateral flow readout enables equipment-free detectionVisual strip results in ~40 minutes (K. Zhang et al. 2024)Demonstrated; cold-chain-free reagent stability still improving
One-pot formats reduce contamination riskSingle-tube RPA + CRISPR/Cas13d detection (D. You et al. 2024)Supported; temperature compatibility between steps can be challenging
Multiplexed detection is achievableDual HPV type detection in single assay (K. Zhang et al. 2024)Demonstrated for 2 targets; scaling to 5+ targets emerging

Open Questions

  • Regulatory approval: No CRISPR diagnostic has received FDA clearance for clinical use. What clinical validation data are required for regulatory approval?
  • Sample preparation: CRISPR diagnostics require nucleic acid extraction from clinical samples. Can simplified extraction (heat lysis, paper-based capture) replace kit-based methods?
  • Quantification: Lateral flow strips provide yes/no results. Can CRISPR diagnostics be made quantitative for viral load monitoring?
  • Cost at scale: Can CRISPR diagnostic reagents (guide RNAs, Cas proteins, isothermal amplification enzymes) be manufactured at the cost points needed for mass deployment in low-resource settings?
  • Referenced Papers

    • [1] Zhang, K. et al. (2024). RPA-CRISPR-Cas Dual Lateral Flow Assay for HPV16 and HPV18. Bioconjugate Chemistry. DOI: 10.1021/acs.bioconjchem.4c00375
    • [2] You, D. et al. (2024). One-pot RPA-CRISPR/EsCas13d platform for JEV detection. Int. J. Biological Macromolecules. DOI: 10.1016/j.ijbiomac.2024.134151
    • [3] Chen, J. et al. (2024). Recent advances in CRISPR/Cas system-based visual detection. Analytical Methods. DOI: 10.1039/d4ay01147c
    • [4] Zhang, H. et al. (2024). RT-ERA-CRISPR/Cas13a lateral flow for TBEV detection. Int. J. Biological Macromolecules. DOI: 10.1016/j.ijbiomac.2024.133720
    • [5] Sun, D. et al. (2025). RT-ERA-CRISPR/Cas12a for duck hepatitis A virus detection. Poultry Science. DOI: 10.1016/j.psj.2025.105316

    References (5)

    Zhang, K., Li, Q., Wang, K., Zhang, Q., Ma, C., Yang, G., et al. (2024). RPA-CRISPR-Cas-Mediated Dual Lateral Flow Assay for the Point-of-Care Testing of HPV16 and HPV18. Bioconjugate Chemistry, 35(11), 1797-1804.
    You, D., Xu, T., Huang, B., Zhu, L., Wu, F., Deng, L., et al. (2024). Rapid, sensitive, and visual detection of swine Japanese encephalitis virus with a one-pot RPA-CRISPR/EsCas13d-based dual readout portable platform. International Journal of Biological Macromolecules, 277, 134151.
    Chen, J., Su, H., Kim, J. H., Liu, L., & Liu, R. (2024). Recent advances in the CRISPR/Cas system-based visual detection method. Analytical Methods, 16(39), 6599-6614.
    Zhang, H., Wang, Y., Chen, C., Xing, W., Xia, W., Fu, W., et al. (2024). A novel rapid visual nucleic acid detection technique for tick-borne encephalitis virus by combining RT-recombinase-aided amplification and CRISPR/Cas13a coupled with a lateral flow dipstick. International Journal of Biological Macromolecules, 275, 133720.
    Sun, D., Zhu, Y., Wang, M., Wang, J., Cheng, W., Li, Z., et al. (2025). A RT-ERA-CRISPR/Cas12a assay for rapid point-of-care duck hepatitis A virus detection. Poultry Science, 104(8), 105316.

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