Journal
ANALYTICAL CHEMISTRY
Volume 93, Issue 38, Pages 12921-12929Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c02228
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Funding
- National Natural Science Foundation of China [22174163]
- Hunan Provincial Science and Technology Plan Project, China [2019TP1001]
- Innovation Driven Project of Central South University [2020CX002]
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A CRISPR-Cas12a-based CTC detection sensor regulated by MDANs and a 3D DNA network formed by RCA products showed improved sensitivity for detecting CTCs. The method was successfully applied to detect CTCs in human blood samples, indicating great potential for clinical CTC-based liquid biopsy.
Although circulating tumor cells (CTCs) have great potential to act as the mini-invasive liquid biopsy cancer biomarker, a rapid and sensitive CTC detection method remains lacking. CRISPR-Cas12a has recently emerged as a promising tool in biosensing applications with the characteristic of fast detection, easy operation, and high sensitivity. Herein, we reported a CRISPR-Cas12a-based CTC detection sensor that is regulated by the multivalent duplexed-aptamer networks (MDANs). MDANs were synthesized on a magnetic bead surface by rolling circle amplification (RCA), which contain multiple duplexed-aptamer units that allow structure switching induced by cell-binding events. The presence of target cells can trigger the release of free activator DNA from the MDANs structure to activate the downstream CRISPR-Cas12a for signal amplification. Furthermore, the 3D DNA network formed by RCA products also provided significantly higher sensitivity than the monovalent aptamer. As a proof-of-concept study, we chose the most widely used sgc8 aptamer that specifically recognizes CCRF-CEM cells to validate the proposed approach. The MDANs-Cas12a system could afford a simple and fast CTC detection workflow with a detection limit of 26 cells mL-1. We also demonstrated that the MDANs-Cas12a could directly detect the CTCs in human blood samples, indicating a great potential of the MDANs-Cas12a in clinical CTC-based liquid biopsy.
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