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Rolling circle amplification for next-generation molecular diagnostics, genome analysis, and spatial transcriptome
Jueun Han1, Yejin Song1, Jihee Lee1
1Department of Bioengineering & Nano-Bioengineering, Research Center for Bio Materials and Process Development, Incheon National University, Incheon 22012, Republic of Korea. e.kim@inu.ac.kr.
Nanoscale
|September 8, 2025
Summary
Rolling circle amplification (RCA) is a powerful isothermal DNA technology for sensitive molecular diagnostics and multi-omics research. Advancements enhance its use in disease detection, genome analysis, and spatial transcriptomics.
Area of Science:
- Molecular Biology
- Biotechnology
- Genomics
Background:
- Rolling circle amplification (RCA) is a robust isothermal DNA amplification method.
- It offers high sensitivity, specificity, and scalability for molecular diagnostics and multi-omics.
- RCA generates long, repetitive DNA sequences with high fidelity.
Purpose of the Study:
- To review the principles, modifications, and applications of RCA.
- To highlight RCA's role in molecular diagnostics, genome analysis, and spatial transcriptomics.
- To discuss challenges and future directions for RCA.
Main Methods:
- Exploration of various RCA formats (solution-phase, solid-phase, hydrogel-based, digital).
- Integration of RCA with CRISPR, biosensors, and nanoparticle amplification.
- Application in extrachromosomal circular DNA sequencing and in situ RNA sequencing.
Main Results:
- RCA enables ultrasensitive detection of nucleic acids, proteins, and cells.
- It provides insights into genome instability, cancer evolution, and cellular heterogeneity.
- RCA facilitates high-resolution gene expression mapping in tissues.
Conclusions:
- RCA is a versatile tool transforming molecular diagnostics and biomedical research.
- Future integration with high-throughput platforms will advance biomarker discovery and multi-omics.
- RCA is poised for a significant role in precision medicine and next-generation sequencing.
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