Epigenetic Differentiation of Monozygotic Twins in Forensic Investigations Using Nanopore Sequencing
Organized by: International Forensic Scientist Awards
Website: forensicscientist.org
13th Edition of Forensic Scientist Awards 28-29 August 2025 | Berlin Germany
Introduction
Monozygotic twins (MZTs), often called identical twins, present a unique challenge in forensic science. Since they share the same nuclear DNA, conventional genetic markers like STRs (Short Tandem Repeats) and SNPs (Single Nucleotide Polymorphisms) cannot distinguish them. This creates major obstacles in forensic investigations, disaster victim identification, and paternity testing.
To overcome this, researchers are now exploring the role of epigenetics, specifically DNA methylation, which reflects subtle differences shaped by environment and lifestyle, even in genetically identical individuals.
Why Epigenetics?
Unlike traditional DNA analysis, epigenetic markers go beyond the genetic code. DNA methylation patterns change over time due to environmental and random (stochastic) influences. These changes make epigenetics an ideal tool to differentiate monozygotic twins in forensic casework.
Nanopore Sequencing: A Breakthrough Technology
Oxford Nanopore sequencing provides:
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Long-read capability – enabling genome-wide methylation profiling
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Single-base resolution – precise detection of methylation changes
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PCR-free analysis – suitable for degraded or trace forensic samples
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High efficiency – with >99.5% alignment and read lengths over 13 kb
These features make nanopore sequencing a promising forensic tool.
Key Findings of the Study
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Analyzed six pairs of MZTs using nanopore sequencing.
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Identified 3,820 differentially methylated loci (DMLs) across the genome.
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DMLs were enriched in metabolic and neural pathways.
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Most were located in intergenic regions (88.03%), with a smaller fraction in promoter regions (1.84%).
These methylation differences serve as stable and heritable biomarkers for distinguishing monozygotic twins.
Forensic Applications
The findings have transformative potential in:
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Criminal investigations – resolving cases where identical twins are suspects.
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Disaster victim identification (DVI) – providing precision in large-scale tragedies.
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Paternity testing – ensuring accurate identification in legal disputes.
Conclusion
This research bridges the gap between epigenetics and forensic science, proving that nanopore-based methylation profiling can successfully differentiate monozygotic twins. By identifying stable epigenetic biomarkers, forensic scientists now have powerful tools to solve cases once thought impossible.
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