Related Experiment Video
Updated: Jan 8, 2026
Clinical Microfluidic Chip Platform for the Isolation of Versatile Circulating Tumor Cells
Published on: October 13, 2023
Simultaneous and sensitive detection of two tumor-related circRNAs by enzyme-free isothermal amplification assisted
Xing Geng1, Shuang Tang1, Jianan Lv1
1School of Chemistry and Molecular Engineering, East China Normal University, 500 Dongchuan Road, Shanghai, 200241, PR China.
Abstract:
Covalently closed single-stranded circular RNAs (circRNAs) are a class of naturally occurring special single-stranded non-coding RNAs with a covalently closed circular structure. CircRNAs have a profound relationship with human cancers and are identified as reliable tumor markers. This study developed an enzyme-free strategy based on catalytic hairpin assembly (CHA) combined with microchip electrophoresis (MCE) to achieve simultaneous detection of circMTO1 and circCDYL. During the CHA process, the well-designed hairpin probes (Hp1 and Hp2) could specifically recognize the back-splicing junction (BSJ) sequence of circRNAs and generate Hp1-Hp2 double-stranded products. Owing to their different numbers of bases, the Hp1-Hp2 double-stranded products corresponding to the two target circRNAs after CHA could be separated and sensitively detected by MCE. Under the optimized conditions, the limits of detection (LOD) of circMTO1 and circCDYL were as low as 2.13 pM and 1.68 pM (S/N = 3), respectively. This approach was also applied to detect circRNAs in HeLa cells, demonstrating excellent applicability and accuracy in biological samples.
Insights
This study introduces an enzyme-free method for detecting cancer-related circular RNAs (circRNAs) using catalytic hairpin assembly and microchip electrophoresis. The novel approach offers sensitive and simultaneous detection of specific circRNAs in biological samples.
Area of Science:
- Biochemistry
- Molecular Biology
- Analytical Chemistry
Background:
- Circular RNAs (circRNAs) are non-coding RNAs with a unique covalently closed structure.
- CircRNAs play significant roles in human cancers and serve as potential tumor markers.
- Accurate and sensitive detection of specific circRNAs is crucial for cancer diagnostics.
Purpose of the Study:
- To develop an enzyme-free strategy for simultaneous detection of circMTO1 and circCDYL.
- To combine Catalytic Hairpin Assembly (CHA) with Microchip Electrophoresis (MCE) for enhanced circRNA analysis.
- To establish a sensitive and accurate method for circRNA detection in biological samples.
Main Methods:
- An enzyme-free strategy utilizing Catalytic Hairpin Assembly (CHA) was employed.
- Well-designed hairpin probes specifically recognized the back-splicing junction (BSJ) of target circRNAs.
- Microchip Electrophoresis (MCE) was used for separation and sensitive detection of CHA products.
Main Results:
- Simultaneous detection of circMTO1 and circCDYL was achieved.
- The method demonstrated high sensitivity with limits of detection as low as 2.13 pM for circMTO1 and 1.68 pM for circCDYL.
- The approach was successfully validated in HeLa cells, showing excellent applicability and accuracy.
Conclusions:
- The developed CHA-MCE method provides a sensitive, enzyme-free, and simultaneous approach for circRNA detection.
- This strategy holds promise for the development of novel diagnostic tools for cancer.
- The method's applicability in biological samples highlights its potential for clinical use.
