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Programmable Ligation-Transcription Circuit-Driven Cascade Amplification Machinery for Multiple Long Noncoding RNAs
Yan Zhang1,2, Xue-Ke Du1, Wen-Jing Liu3
1College of Chemistry, Chemical Engineering and Materials Science, Shandong Normal University, Jinan 250014, China.
Analytical Chemistry
|July 22, 2022
Summary
We developed a sensitive method to detect multiple long noncoding RNAs (lncRNAs) using a ligation-transcription circuit and single-molecule counting. This technique offers high sensitivity for cancer diagnosis and research.
Area of Science:
- Molecular Biology
- Biotechnology
- Cancer Research
Background:
- Accurate measurement of long noncoding RNAs (lncRNAs) is crucial for diagnosing and treating diseases, particularly cancers.
- Existing methods for lncRNA detection face challenges in sensitivity and simultaneous multi-analyte detection.
Purpose of the Study:
- To develop a simple, highly sensitive method for simultaneous detection of multiple lncRNAs.
- To enable single-molecule counting for precise quantification of lncRNAs.
- To apply the method for discriminating lncRNA expression in non-small cell lung cancer (NSCLC) tissues.
Main Methods:
- A programmable ligation-transcription circuit-driven cascade amplification strategy was employed.
- Simultaneous detection of lncRNA HOTAIR and lncRNA MALAT1 was achieved.
- Signal probes were cyclically digested by duplex-specific nuclease, releasing Cy5 and Cy3 molecules for single-molecule counting after magnetic separation.
Main Results:
- The assay demonstrated extremely high sensitivity with limits of detection (LOD) of 0.043 aM for lncRNA HOTAIR and 0.126 aM for lncRNA MALAT1.
- The method allows for simultaneous measurement of multiple endogenous lncRNAs at the single-cell level.
- Significant differences in lncRNA expression were observed between NSCLC tumor tissues and adjacent healthy tissues.
Conclusions:
- This novel method provides a highly sensitive and specific platform for simultaneous lncRNA detection.
- The technique holds significant promise for clinical diagnosis of cancers like NSCLC.
- It offers a valuable tool for advancing biomedical research and understanding lncRNA functions.
