Forensic Science International-Genetics最新文献

{"title":"ForSpEC: A compact forensic epigenetic age clock for sperm cells with cross‑platform validation","authors":"Aleksandra Pisarek-Pacek ,&nbsp;Joanna Rudnicka ,&nbsp;Rezvan Noroozi ,&nbsp;Bożena Wysocka ,&nbsp;Aleksander Masny ,&nbsp;Magdalena Spólnicka ,&nbsp;Andrzej Ossowski ,&nbsp;Aneta Sitek ,&nbsp;Aneta Macur ,&nbsp;Jarosław Janeczko ,&nbsp;Gabriela Kania ,&nbsp;Marta Sikora-Polaczek ,&nbsp;Wojciech Branicki ,&nbsp;Ewelina Pośpiech","doi":"10.1016/j.fsigen.2026.103426","DOIUrl":"10.1016/j.fsigen.2026.103426","url":null,"abstract":"<div><div>Expanding the use of forensic DNA intelligence requires the development of accurate DNA-based predictive models. The primary challenge in achieving a highly accurate epigenetic age prediction method in sperm lies in the unique biological function of male reproductive cells. In this study, we independently evaluated the Germ Line Age (GLA) calculator, one of the most comprehensive tools for predicting sperm age to date. We then set out to define a streamlined, well-curated set of sperm cell-specific markers suitable for forensic applications, while providing an integrated laboratory tool for targeted methylation profiling using bisulfite amplicon sequencing with Ion AmpliSeq™ technology. DNA was extracted from 212 sperm samples, genome-wide DNA methylation was measured using Infinium MethylationEPIC (EPIC) arrays and methylation beta values were extracted for 260 literature-reported CpG markers. Application of the GLA model to this sperm cell dataset resulted in a mean absolute error (MAE) of 2.82 years. As a next step, predictive analysis based on 147 training set samples identified a subset of eight optimal CpG markers for model building. Due to technical constraints encountered during Ion AmpliSeq™ assay optimization, the final model was refined to include seven CpGs, yielding the ForSpEC (<u>For</u>ensic <u>Sp</u>erm <u>E</u>pigenetic <u>C</u>lock) model. ForSpEC predicted age with an MAE of 3.13 years in the 65 test samples. When the EPIC-trained ForSpEC model was applied to Ion AmpliSeq™ data (N = 20), age prediction accuracy remained high, with an MAE of 3.76 years. For comparison, existing predictors were retrained using EPIC data, demonstrating that the seven CpGs comprising ForSpEC rank among the most accurate predictors of sperm epigenetic age. The model was further validated using data from publicly available repositories and the impact of cellular composition on prediction accuracy was additionally addressed by comparing DNA samples isolated from sperm cells with whole semen samples. Overall, this study advances sperm-based epigenetic age estimation by introducing ForSpEC, a highly accurate forensic epigenetic clock that relies on just seven CpG markers. This new method exhibits only a modest reduction in accuracy compared to the GLA model, which relies on 261 CpGs. Moreover, the analyses underscore the high utility of Ion AmpliSeq™ technology for targeted DNA methylation analysis in forensic applications.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103426"},"PeriodicalIF":3.1,"publicationDate":"2026-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing forensic casework interpretation through DNA methylation-based analysis: A case study of pooled blood samples","authors":"Jeong Min Lee ,&nbsp;Sang Un Park ,&nbsp;Bo Min Kim ,&nbsp;Hanbyeol Kim ,&nbsp;Hwan Young Lee","doi":"10.1016/j.fsigen.2026.103424","DOIUrl":"10.1016/j.fsigen.2026.103424","url":null,"abstract":"<div><div>In forensic casework, the meticulous investigation of unidentified biological material is critical for developing decisive investigative leads. Integrated genetic and epigenetic profiling allows for the reconstruction of incident scenarios and the assessment of criminal involvement probability. Specifically, DNA methylation-based analyses facilitate the determination of cellular composition, donor age, and smoking status, progressively delineating probable incident circumstances and converging on donor identification. We report a complex case involving a large volume of pooled blood discovered on an apartment staircase half-landing with an unknown donor identity. The investigation was refined using multiple genetic and epigenetic assays, including STR typing, Y-haplotyping, SNaPshot-based body fluid identification, and array-based DNA methylation profiling. Two samples collected at the scene—one presumed blood and one unknown—were confirmed to originate from the same donor: a Korean male belonging to Y-haplogroup O3a2. SNaPshot analysis indicated the presumed blood sample was a blood–saliva mixture, while the unknown sample remained unassigned to any specific fluid type. Array-based cell/tissue deconvolution and unsupervised clustering analysis further revealed that the presumed blood sample was predominantly composed of whole blood with a minor contribution from upper gastrointestinal tissue. Integrating the results of body fluid identification with cell-proportion deconvolution supported the interpretation of hematemesis (vomiting blood), thereby favoring a medical scenario over criminal involvement. The unknown sample exhibited methylation patterns distinct from all candidate tissue groups, possibly reflecting sample quality limitations or tissue mixture. Chronological epigenetic clock analyses estimated the donor’s age to be middle-aged (approximately 50–60 years), while smoking status prediction classified the donor as a current smoker. Despite suboptimal sample quality and the complete absence of contextual information, this comprehensive analysis provided detailed donor profiling and incident reconstruction. This case exemplifies the power of integrated genetic and epigenetic methodologies—particularly tissue-of-origin profiling—to generate investigative leads, facilitate the inference of incident circumstances, and provide substantive contributions to forensic investigations.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103424"},"PeriodicalIF":3.1,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145977150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cell-type-dependent expression of a somatic tri-allelic pattern at D18S51 and Penta D: The statistical interpretation of forensic evidence in sexual assault investigations","authors":"A. Sala ,&nbsp;M. Caputo ,&nbsp;C. Schatz ,&nbsp;D. Corach ,&nbsp;T. Egeland ,&nbsp;F. Marsico","doi":"10.1016/j.fsigen.2026.103425","DOIUrl":"10.1016/j.fsigen.2026.103425","url":null,"abstract":"<div><div>Tri-allelic patterns, though rare, present a significant interpretive challenge in forensic genetics. Their misinterpretation as a DNA mixture can lead to flawed statistical assessment, particularly for likelihood ratio (LR) calculations. However, when correctly identified, a tri-allelic pattern can serve as a powerful identifying marker, increasing the evidential value of a match between a POI (person of interest) and forensic evidence. This case report analyses a sexual assault investigation involving the analysis of cloth fragments, subungual, anal, vulvar, and vaginal swabs, alongside reference samples from the victim and suspect. The POI’s buccal reference sample exhibited a tri-allelic pattern at the D18S51 locus (alleles: 16, 17, 18) and a substantial peak height imbalance (PHR = 0.16) at Penta D. We found that this tri-allelic pattern corresponds to a Type 1, an early embryonic somatic mutation. While some evidence items yielded mixed profiles of the victim and suspect, others including a cloth fragment, two subungual swabs, and an anal swab revealed a single-source male profile matching the POI. Critically, the tri-allelic pattern was observed across multiple evidence types, but the relative proportion of the three alleles (the intra-locus ratio) varied systematically depending on the tissue origin of the sample. This finding suggests cell-type-dependent genetic makeup of the somatic mutation, a phenomenon known as mosaicism. We discuss the characteristics of this pattern, postulate its embryonic origin, and propose a framework for its statistical incorporation into the LR. This case underscores that recognition of the biological mechanism is essential to avoid misinterpretation and can provide stronger evidence of genetic identification in forensic casework. The freely available app LRtri, <span><span>https://francomarsico.shinyapps.io/LRtri/</span><svg><path></path></svg></span>, can be used to explore the uncertainty in the LR.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103425"},"PeriodicalIF":3.1,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145977149","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantitative trait loci as indicators of potential susceptibility to allele-specific dropout for forensic RNA markers","authors":"M. van den Berge,&nbsp;T. Sijen","doi":"10.1016/j.fsigen.2026.103423","DOIUrl":"10.1016/j.fsigen.2026.103423","url":null,"abstract":"<div><div>In recent years, the potential to associate donor (from DNA results) with specific cell types (based on RNA analysis) using single nucleotide polymorphisms (SNPs) has been explored and recognized as a major advancement in forensic RNA typing. This method relies on SNPs in transcribed regions of body fluid-specific mRNA to link a cell type to a specific donor, whose presence is inferred from conventional forensic DNA typing. While the approach is promising, discrepancies frequently arise between the DNA and RNA-SNP data, primarily due to allelic dropout of RNA alleles. Understanding the mechanism underlying allele-specific expression is essential for advancing this method towards casework implementation. To address this, the role of quantitative trait loci (QTLs) as a potential cause of allele-specific expression (ASE) was investigated. Findings show how QTLs help explain the previously unresolved discrepancies and how QTL analysis can serve as a predictor of a gene’s susceptibility to ASE. Additionally, guidelines are provided for handling genes with increased risks of ASE, aiding the implementation of DNA and RNA-SNP-based donor-to-cell-type association in forensic casework.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103423"},"PeriodicalIF":3.1,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145977151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Using DNA to restore the identity and dignity of Richmond’s ancestors: The East Marshall Street Well Project","authors":"Filipa Simão ,&nbsp;Sierra Laveroni ,&nbsp;Michelle Woo ,&nbsp;Daniela Frausto ,&nbsp;Amber Mundy ,&nbsp;Martin MacStudy ,&nbsp;Christopher Twene ,&nbsp;Andrea Malchow ,&nbsp;Andrea Sanjurjo ,&nbsp;Baneshwar Singh ,&nbsp;Tal Simmons","doi":"10.1016/j.fsigen.2026.103422","DOIUrl":"10.1016/j.fsigen.2026.103422","url":null,"abstract":"<div><div>In 1994, a 19th-century well containing commingled skeletal remains was discovered in Richmond, Virginia. These remains, primarily stolen from African American burial grounds, had been used at the Medical College of Virginia for dissection practices. The East Marshall Street Well Project was established in 2013 to foster community discussions on this discovery. Following the consultation process, the Family Representative Council (FRC) was formed to represent the descendant community of those found in the well and guide respectful memorialization and reburial efforts. Genetic and anthropological research was approved, aiming to reassociate the commingled remains into individuals and to provide insights into their ancestral origins. Through DNA analysis, bones belonging to 33 individuals were reassociated. Although degradation and commingling limited full reassociation, 36 unique INNUL profiles were identified, establishing a minimum of 36 adult individuals. Mitochondrial DNA haplotypes from the juvenile remains indicated the presence of at least six juveniles. Ancestry-informative markers confirmed predominantly African heritage, with mitochondrial DNA and Y-chromosome data linking the African ancestors of the EMSW individuals to Central-West Africa. This aligns with documented patterns of displacement during the transatlantic slave trade. Anthropological analysis of skeletal biography and postmortem skeletal markers provided insight into the heavy labor endured by these individuals during their lives and the disregard for their bodies after death. This study is crucial for ensuring respectful burial and proper memorialization, honoring the individuals’ personhood, dignity, and historical significance. By shedding light on their origins and experiences, it helps the descendant community address past injustices.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103422"},"PeriodicalIF":3.1,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145977148","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the efficiency of different sets of microhaplotypes for first to third degree kinships","authors":"Xiao-Ye Jin ,&nbsp;Li-Shuai Tan ,&nbsp;Zhi Chen ,&nbsp;Qing-Zhen Meng ,&nbsp;Yao-Sen Feng ,&nbsp;Chi Zhang ,&nbsp;Ke-Lai Kang ,&nbsp;Jie Zhao ,&nbsp;Jing Sun ,&nbsp;Yu-Heng Wu ,&nbsp;Shi-Yao Sun ,&nbsp;Bofeng Zhu ,&nbsp;Jian Ye ,&nbsp;Le Wang","doi":"10.1016/j.fsigen.2026.103420","DOIUrl":"10.1016/j.fsigen.2026.103420","url":null,"abstract":"<div><div>Microhaplotypes (MHs) are short, multi-allelic genetic markers that combine high genomic abundance with low mutation rates and the absence of stutter artefacts in PCR. These properties make them promising genetic markers for resolving complex kinships in forensic genetics. In this study, 202 MHs were genotyped in 181 family samples. By integrating empirical genotyping with extensive in-silico simulations, we evaluated the ability of these 202 MHs to identify first-, second-, and third-degree relationships from unrelated pairs. Additionally, four previously selected sets of MHs (190, 301, 337 and 703 MHs) were re-examined to determine their respective effectiveness across these kinship classes. The 202 MHs reliably distinguished first-degree relatives from unrelated individuals and other types of kinships. In addition, these 202 MHs could also discriminate second-degree relatives from unrelated individuals with the accuracy &gt; 0.99. In contrast, the panel’s performance declined markedly for third-degree relationships, and obtained accuracy was less than 0.5 when analytical threshold of cumulative likelihood ratio was set as 10⁴ and 10⁻⁴. The four alternative panels (190, 301, 337 and 703 MHs) exhibited similarly high effectiveness for first- and second-degree relationships with the accuracy &gt; 0.99. For third-degree relationships, accuracy increased monotonically with the number of microhaplotypes. Specifically, the 337-MH and 703-MH sets correctly assigned the majority of third-degree pairs regardless of the threshold adopted, indicating their suitability for third-degree kinship testing. In summary, the 202-MH panel could serve as a highly efficient tool for first- and second-degree kinship testing in forensic practice. In addition, panels containing 337 or 703 MHs are recommended for third-degree analyses. Future work should further improve performance by integrating MHs with other types of genetic markers to address intricated kinship analysis.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103420"},"PeriodicalIF":3.1,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145938493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Long non-coding RNAs in human peripheral blood as biomarkers for age prediction","authors":"Anqi Chen ,&nbsp;Yujia Xuan ,&nbsp;Fan Yang ,&nbsp;Yanan Liu ,&nbsp;Jinyuan Zhao ,&nbsp;Mengxiao Liao ,&nbsp;Yanfang Lu ,&nbsp;Yu Xing ,&nbsp;Suhua Zhang ,&nbsp;Chengtao Li","doi":"10.1016/j.fsigen.2025.103419","DOIUrl":"10.1016/j.fsigen.2025.103419","url":null,"abstract":"<div><div>Chronological age is a critical biological trait with substantial relevance in forensic investigations. Growing evidence highlights the role of long non-coding RNAs (lncRNAs) in aging, but the application in age prediction remains largely unexplored. To evaluate the potential of lncRNAs for age prediction, we analyzed RNA-seq data from peripheral blood samples of Han Chinese individuals. Differential expression analysis identified a series of differentially expressed lncRNAs (DElncRNAs) among multiple age group comparisons, with the most significant expression changes observed between the youngest (18–27 years) and older (68–77 years) age groups. GO and KEGG enrichment analysis of target genes indicated that these lncRNAs were involved in mitochondrial function, telomere maintenance, and pathways related to neurodegenerative disease. Additionally, Spearman’s correlation analysis further identified 20 lncRNAs significantly associated with age as candidate markers, which were subsequently used to construct age prediction models using six machine learning algorithms. The XGBoost model achieved the best performance, with a mean absolute error (MAE) of 8.04 years (R² = 0.66) in the test cohort. External validation was conducted using two independent datasets (GSE124326 and GSE177044). The XGBoost and LightGBM models consistently outperformed the other models, with MAEs ranging from 7.99 to 8.21 years. These findings demonstrated the feasibility of lncRNA-based age prediction, providing novel biomarkers and methodological insights for forensic age estimation.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103419"},"PeriodicalIF":3.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145938492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of DNA recovery from improvised explosive device components after detonations","authors":"Ghassan Ali Salih ,&nbsp;Martina Nilsson ,&nbsp;Moa Lidén ,&nbsp;Marie Allen","doi":"10.1016/j.fsigen.2025.103418","DOIUrl":"10.1016/j.fsigen.2025.103418","url":null,"abstract":"<div><div>Improvised explosive devices (IEDs) are commonly utilised by terrorist and criminal groups, and can be constructed using inexpensive and readily accessible materials. Despite the challenges posed by detonation, the DNA of those who made or placed the device may be recovered from the components of detonated IEDs. However, the recovered DNA is often scarce, degraded, damaged, and in the form of both cell-bound and cell-free DNA from, for example, skin contact. This study examines the recovery and the success of downstream analysis of DNA from various biological sources, including cell-bound blood and touch DNA and extracted cell-free DNA. DNA from different sources was deposited on different IED components with different surface materials. The IEDs were detonated using 40 g, 80 g, or 125 g of dynamite and DNA was sampled and analysed using real-time qPCR, STR analysis and mtDNA sequencing. The results indicated that multiple factors, including the type of DNA from different biological sources, dynamite weight, component type, DNA location and surface material, influenced the rate of successful genotyping. It was also demonstrated that the DNA quantities, and the success in STR profiling and mtDNA amplification were highly correlated for both cell-bound DNA and cell-free DNA. Moreover, sampling from internal phone components and PVC surfaces demonstrated higher DNA-typing success rates than DNA on external phone surfaces or metal-based components (SIM card holder, electrical wires, and circuit boards). DNA from blood revealed the highest rates of successful STR genotyping results across most detonated IED components, even for the highest weights of dynamite tested. Furthermore, mtDNA sequencing demonstrated a high success rate for low-template samples, even after the use of high amounts of explosive weight. A majority of DNA traces found on detonated IEDs are likely to consist of touch DNA from the production of the device. Here, the touch DNA samples on 9 V batteries yielded more DNA and higher success in DNA profiling than DNA on toggle switches and metal-containing electrical wires. These findings provide valuable insights into practical strategies for DNA recovery and analysis in IED-related forensic investigations.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103418"},"PeriodicalIF":3.1,"publicationDate":"2025-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145938491","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Target RNA sequencing system for identification of body fluid contributors in mixtures","authors":"Huan Yu,&nbsp;Jiaxin Ji,&nbsp;Jiayan Li,&nbsp;Jiaqi Wang,&nbsp;Zidong Liu,&nbsp;Xiaoxiao Wang,&nbsp;Yiwei Guo,&nbsp;Liangze Li,&nbsp;Jinding Liu,&nbsp;Zeqin Li,&nbsp;Gengqian Zhang","doi":"10.1016/j.fsigen.2025.103416","DOIUrl":"10.1016/j.fsigen.2025.103416","url":null,"abstract":"<div><div>The cSNPs within body fluid-specific RNA molecules have emerged as a promising biomarker for body fluid identification and identifying body fluid donors in mixture stains. Unlike traditional DNA analysis, which is restricted to individualizing a forensic stain at a sub-source level, RNA-SNP profiling enables direct association of the body fluid with the suspect. However, the number of available RNA-SNPs is still not enough to achieve a satisfactory discriminatory power. To explore novel SNPs for RNA-based genotyping, we screened out a series of potential RNA markers and developed a target RNA sequencing assay comprising 135 RNA-SNPs across 42 body fluid genes to identify body fluid contributors in the mixture. The assessment of specificity was performed on single-source samples, and the results exhibited a clear separation between five body fluid types. The assay also showed complete genotype concordance between gDNA and cDNA, and had a capacity to analyze aged stains. In the artificial RNA mixtures consisting of 2–5 body fluid components and 2-component mock case mixtures, all body fluid components could be successfully detected, and most of the genotypes could be correctly associated with their body fluid contributors, indicating excellent discriminative capacity of our panel for mixture deconvolution. The panel presented a cumulative discriminatory power ranging from 0.987325116 (saliva) to 0.999993412 (venous blood), and could serve as an effective complementary tool to improve discriminatory power when combined with other currently available RNA-SNP panels.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103416"},"PeriodicalIF":3.1,"publicationDate":"2025-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145866411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Challenges in mRNA-based time since deposition estimation: A multiplex MPS primer panel investigation","authors":"Nadescha Viviane Hänggi ,&nbsp;Melisa Walliser ,&nbsp;Ane Elida Fonneløp ,&nbsp;Robert Lagacé ,&nbsp;Erin Hanson ,&nbsp;Jack Ballantyne ,&nbsp;Cordula Haas","doi":"10.1016/j.fsigen.2025.103415","DOIUrl":"10.1016/j.fsigen.2025.103415","url":null,"abstract":"<div><div>Estimating the time since deposition (TsD) of biological stains can provide crucial information for crime scene investigations, but remains a challenging task. In this study, we investigated mRNA degradation patterns across four forensically relevant body fluids - blood, semen, menstrual blood, and vaginal secretion - using a custom-designed mRNA MPS primer panel. The primer panel included candidate mRNA markers identified by RNA-Seq for TsD estimation in multiple body fluids. It was designed to assess mRNA degradation patterns based on three aspects: (1) individual mRNA degradation trends, (2) intra-transcript stability, examining 5′- versus 3’-end degradation, and (3) inter-transcript stability, based on relative degradation between two targets. The results revealed high inter-individual variability, especially for low-abundance transcripts. Some mRNA markers showed degradation patterns that correlated with TsD. However, inconsistencies in the degradation patterns observed across the analyzed time series highlight the challenges associated with primer panel optimization, RNA extraction methods, and reproducibility across samples. Our results suggest that, while mRNA degradation remains a promising strategy for TsD estimation, its forensic application requires careful marker selection and validation across independent data sets, to account for methodological and biological variability.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103415"},"PeriodicalIF":3.1,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145919497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}