ArticleInternational journal of legal medicine2026
CircRNA profiles from one-tube multiplex probe-based qPCR can distinguish between monozygotic twins.
Article in International journal of legal medicine, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Monozygotic twins (MZTs) pose a considerable challenge in personal identification due to their nearly identical genomic DNA sequences. Emerging evidence suggests that epigenetic markers, particularly non-coding RNAs (ncRNAs), may provide a promising strategy to discriminate MZTs. Circular RNAs (circRNAs) feature high stability and tissue-specific expression and have shown considerable potential in forensic applications such as body fluid identification and age estimation. In this study, we performed whole-transcriptome circRNA sequencing on peripheral blood samples from five MZT pairs to screen for differentially expressed circRNAs (DEcircRNAs). A total of 13,191 circRNAs were identified, and six DEcircRNAs (hsa_circ_0018054, hsa_circ_0003413, hsa_circ_0067717, hsa_circ_0003941, hsa_circ_0107772, and hsa_circ_0058520) were confirmed by real-time quantitative polymerase chain reaction (RT-qPCR) in an expanded cohort of 12 MZT pairs. Furthermore, these markers exhibited high stability in dried bloodstains stored for up to 180 days. Additionally, we developed a novel one-tube multiplex probe-based qPCR system for simultaneous detection of three circRNAs (hsa_circ_0003413, hsa_circ_0067717, and hsa_circ_0058520). This system enabled sensitive detection with a minimum requirement of just 4 ng of total RNA. Collectively, our study demonstrates that circRNAs are differentially expressed in MZTs, representing a novel epigenetic biomarker set for forensic individual identification. Our findings support the potential of circRNAs as stable and informative biomarkers to discriminate monozygotic twins in forensic practice.
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