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A programmable and sensitive CRISPR/Cas12a-based MicroRNA detection platform combined with hybridization chain
Hai-Yan Jia1, Hong-Li Zhao1, Ting Wang1
1Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
Biosensors & Bioelectronics
|May 23, 2022
Summary
This study introduces a novel CRISPR/Cas12a and hybridization chain reaction (HCR) platform for sensitive microRNA (miRNA) detection. The method enhances diagnostic capabilities for cancer by enabling versatile miRNA analysis without crRNA redesign.
Area of Science:
- Molecular Biology
- Biotechnology
- Cancer Diagnostics
Background:
- MicroRNAs (miRNAs) are crucial biomarkers for cancer diagnosis and prognosis.
- Existing CRISPR-based miRNA detection methods necessitate custom crRNA synthesis for each target, limiting broad applicability.
- There is a continuous need for sensitive and versatile miRNA detection technologies.
Purpose of the Study:
- To develop a versatile and highly sensitive miRNA detection platform.
- To overcome the limitations of current CRISPR-based methods by eliminating the need for target-specific crRNA synthesis.
- To establish a practical tool for cancer diagnosis and therapy monitoring.
Main Methods:
- A novel platform integrating the CRISPR/Cas12a system with a hybridization chain reaction (HCR) circuit was designed.
- The HCR circuit acts as a signal transducer, amplifying the miRNA signal by generating multiple DNA duplexes.
- These DNA duplexes activate the Cas12a trans-cleavage activity for enhanced signal amplification.
Main Results:
- The platform demonstrated sensitive detection of different miRNAs without requiring changes to the crRNA spacer sequence.
- Signal amplification was achieved through the synergistic action of HCR and Cas12a trans-cleavage.
- The proposed method showed consistency with RT-qPCR for miRNA detection in various cell lines, validating its practicality.
Conclusions:
- The developed CRISPR/Cas12a-HCR platform offers a versatile and sensitive approach for miRNA detection.
- This technology eliminates the need for specific crRNA synthesis, simplifying the detection process.
- The platform shows significant potential for clinical applications in cancer diagnosis and therapy monitoring.
