Journal
ACS SENSORS
Volume 5, Issue 7, Pages 2239-2246Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.0c00944
Keywords
extracellular vesicles; CRISPR-Cas12a; trans-cleavage; CD63 aptamer; allosteric probe; wash-free
Funding
- Fundamental Research Funds for the Central Universities [2019CDYGZD005, 2020CDJYGRH-YJ05]
- National Natural Science Foundation of China [81871733, 81572079]
- postgraduate Supervisor Team Funds for the Chongqing Education Commission
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Extracellular vesicles (EVs) are emerging as promising biomarkers for cancer diagnosis and therapy. Recognizing low-abundance EVs from clinical samples in an easy-to-operate way is highly desired but remains a challenge. Herein, we established an allosteric probe-initiated dual cycle amplification-assisted CRISPR-Cas12a (AID Cas) platform for sensitive detection of EVs in a wash-free way. In AID-Cas, the allosteric probe can specifically recognize and bind with target EVs and thus initiate the following dual-cycle amplification. Subsequently, the amplified products were transcribed to generate numerous single-stranded RIVAs, which could work as crRNA to trigger the trans cleavage of CRISPR-Cas12a. Consequently, the proposed approach achieved a good linear response to extracted EVs in a concentration range from 10(2) to 10(6) particles/mu L. Because of its high sensitivity, together with its wash-free convenience, the proposed strategy could have promising clinical potentials for early diagnosis of cancers.
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