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Hybridization chain reaction circuit controller: CRISPR/Cas12a conversion amplifier for miRNA-21 sensitive detection
Keyi Long1, Gaihua Cao1, Yue Qiu1
1Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China.
Talanta
|September 1, 2023
Summary
We developed a novel HCR-Cas12a controller for ultra-sensitive microRNA (miRNA) detection. This method enables accurate diagnosis of diseases by detecting low-abundance miRNA-21 (miR-21) with high sensitivity.
Area of Science:
- Biochemistry
- Molecular Biology
- Diagnostics
Background:
- MicroRNA (miRNA) detection is vital for disease diagnosis but challenging due to short length and low abundance.
- Existing methods often require complex equipment and lack sensitivity for low-concentration targets.
Purpose of the Study:
- To develop a sensitive and accessible method for detecting microRNA-21 (miR-21).
- To establish a CRISPR/Cas12a-based system for enhanced miRNA detection.
Main Methods:
- Designed a hybridization chain reaction (HCR) circuit controller to initiate a CRISPR/Cas12a amplification system.
- Utilized Cas12a's RNase activity for self-maturation of pre-crRNA and trans-cleavage activity for signal amplification.
- Converted biological signals from miR-21 presence into fluorescent signals.
Main Results:
- Achieved ultra-sensitive detection of miR-21 with a wide linear range (1 fM - 100 nM) and a limit of detection as low as 75.4 aM.
- Demonstrated sensitivity an order of magnitude higher than standard methods.
- HCR formation occurred at room temperature, eliminating the need for thermal cyclers.
Conclusions:
- The HCR-Cas12a controller offers a highly sensitive and rapid method for miRNA detection.
- The system is suitable for low-resource settings due to its simplicity and lack of requirement for expensive instruments.
- Provides a foundation for sensitive detection of low-concentration miRNAs, aiding disease diagnosis.
