Theranostics 2020; 10(22):10262-10273. doi:10.7150/thno.49047 This issue Cite

Research Paper

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

Shan Xing1,2#, Zedong Lu1#, Qi Huang1#, Huilan Li1, Yu Wang1, Yanzhen Lai1,3, Yi He1, Min Deng4✉, Wanli Liu1✉

1. Department of Clinical Laboratory, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangdong Key Laboratory of Nasopharyngeal Carcinoma Diagnosis and Therapy, Sun Yat-sen University Cancer Center, 651 Dongfeng Road East, Guangzhou 510060, P. R. China.
2. School of Biomedical Engineering, Sun Yat-sen University, No. 132 Waihuandong Road, University Town, Guangzhou 510006, PR China.
3. Heyuan People's Hospital, Heyuan, China.
4. Affiliated Cancer Hospital & Institute of Guangzhou Medical University, No.78, Hengzhigang Road, Guangzhou 510095, P. R. China.
#These authors contributed equally to this work.

Citation:
Xing S, Lu Z, Huang Q, Li H, Wang Y, Lai Y, He Y, Deng M, Liu W. An ultrasensitive hybridization chain reaction-amplified CRISPR-Cas12a aptasensor for extracellular vesicle surface protein quantification. Theranostics 2020; 10(22):10262-10273. doi:10.7150/thno.49047. https://www.thno.org/v10p10262.htm
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Abstract

Graphic abstract

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.

Methods: 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.

Results: 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 102 particles/µL, which is at least 104-fold more sensitive than aptamer-ELISA and 102-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 nucleolin+ TEVs for nasopharyngeal carcinoma cancer (NPC) diagnosis and PD-L1+ TEVs for therapeutic monitoring.

Conclusion: 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.

Keywords: Tumor-derived exosomes, CRISPR-Cas12a, Hybridization chain reaction, Aptamer, Fluorescence


Citation styles

APA
Xing, S., Lu, Z., Huang, Q., Li, H., Wang, Y., Lai, Y., He, Y., Deng, M., Liu, W. (2020). An ultrasensitive hybridization chain reaction-amplified CRISPR-Cas12a aptasensor for extracellular vesicle surface protein quantification. Theranostics, 10(22), 10262-10273. https://doi.org/10.7150/thno.49047.

ACS
Xing, S.; Lu, Z.; Huang, Q.; Li, H.; Wang, Y.; Lai, Y.; He, Y.; Deng, M.; Liu, W. An ultrasensitive hybridization chain reaction-amplified CRISPR-Cas12a aptasensor for extracellular vesicle surface protein quantification. Theranostics 2020, 10 (22), 10262-10273. DOI: 10.7150/thno.49047.

NLM
Xing S, Lu Z, Huang Q, Li H, Wang Y, Lai Y, He Y, Deng M, Liu W. An ultrasensitive hybridization chain reaction-amplified CRISPR-Cas12a aptasensor for extracellular vesicle surface protein quantification. Theranostics 2020; 10(22):10262-10273. doi:10.7150/thno.49047. https://www.thno.org/v10p10262.htm

CSE
Xing S, Lu Z, Huang Q, Li H, Wang Y, Lai Y, He Y, Deng M, Liu W. 2020. An ultrasensitive hybridization chain reaction-amplified CRISPR-Cas12a aptasensor for extracellular vesicle surface protein quantification. Theranostics. 10(22):10262-10273.

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