An ultrasensitive hybridization chain reaction-amplified CRISPR-Cas12a aptasensor for extracellular vesicle surface protein quantification

Sep 15, 2020Theranostics

Highly sensitive CRISPR-based sensor for measuring proteins on small cell-released particles

AI simplified

CRISPR Gene Editing on OpenScience ↗PubMed ↗DOI ↗

Abstract

Limit of detection values of 10 particles/µL for tumor-derived extracellular vesicle proteins were achieved, demonstrating high sensitivity.

The apta-HCR-CRISPR assay enables direct and high-sensitivity detection of tumor-derived extracellular vesicle proteins.
Detection of the protein markers nucleolin and programmed death ligand 1 (PD-L1) was successfully verified.
The assay is at least 10-fold more sensitive than existing methods such as aptamer-ELISA and apta-HCR-ELISA.
Clinical analysis of circulating tumor-derived extracellular vesicles from serum samples indicates potential for cancer diagnosis and therapeutic monitoring.

AI simplified

Tumor-derived extracellular vesicle (TEV) protein biomarkers facilitate cancer diagnosis and prognostic evaluations. However, the lack of reliable and convenient quantitative methods for evaluating TEV proteins prevents their clinical application.Here, based on dual amplification of hybridization chain reaction (HCR) and CRISPR-Cas12a, we developed the apta-HCR-CRISPR assay for direct high-sensitivity detection of TEV proteins. The TEV protein-targeted aptamer was amplified by HCR to produce a long-repeated sequence comprising multiple CRISPR RNA (crRNA) targetable barcodes, and the signals were further amplified by CRISPR-Cas12a collateral cleavage activities, resulting in a fluorescence signal.The established strategy was verified by detecting the TEV protein markers nucleolin and programmed death ligand 1 (PD-L1). Both achieved limit of detection (LOD) values as low as 10particles/µL, which is at least 10-fold more sensitive than aptamer-ELISA and 10-fold more sensitive than apta-HCR-ELISA. We directly applied our assay to a clinical analysis of circulating TEVs from 50 µL of serum, revealing potential applications of nucleolinTEVs for nasopharyngeal carcinoma cancer (NPC) diagnosis and PD-L1TEVs for therapeutic monitoring.The platform was simple and easy to operate, and this approach should be useful for the highly sensitive and versatile quantification of TEV proteins in clinical samples. Methods: Results: Conclusion: 2 4 2 + +

Full Text

We can’t show the full text here under this license. Use the link below to read it at the source.

View full text ↗

An ultrasensitive hybridization chain reaction-amplified CRISPR-Cas12a aptasensor for extracellular vesicle surface protein quantification

Abstract

Full Text

You found one interesting study. We’ll send the next 7.

what lands in your inbox each week:

Recent issues from the crispr gene editing brief

Abstract

Full Text

Related papers

You found one interesting study. We’ll send the next 7.

what lands in your inbox each week:

Recent issues from the crispr gene editing brief