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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
PubMed
Summary
This summary is machine-generated.

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.

Keywords:
CRISPR/CasDiseases diagnoseHybridization chain reaction (HCR)MicroRNANucleic acid detection

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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.