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Multivalent Duplexed-Aptamer Networks Regulated a CRISPR-Cas12a System for Circulating Tumor Cell Detection
Zhengxian Lv1, Qiuquan Wang1, Minghui Yang2
1Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis and Instrumentation, College of Chemistry and Chemical Engineering and State Key Lab of Marine Environmental Science, Xiamen University, Xiamen 361005, China.
A new CRISPR-Cas12a sensor using multivalent duplexed-aptamer networks (MDANs) offers rapid and sensitive detection of circulating tumor cells (CTCs). This technology shows promise for non-invasive liquid biopsy in cancer diagnostics.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Circulating tumor cells (CTCs) are valuable biomarkers for liquid biopsy but require sensitive detection methods.
- CRISPR-Cas12a offers fast, sensitive, and user-friendly biosensing capabilities.
- Existing CTC detection methods often lack the required speed and sensitivity for clinical application.
Purpose of the Study:
- To develop a rapid and highly sensitive sensor for detecting circulating tumor cells (CTCs).
- To leverage CRISPR-Cas12a technology combined with a novel aptamer-based signal amplification system.
- To validate the sensor's performance in detecting cancer cells in blood samples.
Main Methods:
- Synthesis of multivalent duplexed-aptamer networks (MDANs) on magnetic beads via rolling circle amplification (RCA).
- Utilizing cell-binding events to trigger the release of activator DNA, activating CRISPR-Cas12a.
- Employing the sgc8 aptamer for specific recognition of CCRF-CEM cells in a proof-of-concept study.
Main Results:
- The MDANs-Cas12a system demonstrated a low detection limit of 26 cells/mL for CTCs.
- The 3D DNA network structure enhanced sensitivity compared to monovalent aptamers.
- Successful direct detection of CTCs in human blood samples was achieved.
Conclusions:
- The MDANs-Cas12a sensor provides a simple, fast, and sensitive method for CTC detection.
- This approach holds significant potential for clinical applications in CTC-based liquid biopsy.
- The developed sensor addresses the need for improved non-invasive cancer biomarker detection.
