- •Two sex-biased genes are found suitable to infer sex in RNA assays.
- •XIST is female-expressed from its function in X-chromosome inactivation.
- •RPS4Y1 is Y-chromosome-specific and therefore male-expressed.
- •Both markers are incorporated in two multiplexes targeting body fluids or organs.
- •These assays were validated for casework.
In forensics, DNA profiling is used for the identification of the donor of a trace, while messenger RNA (mRNA) profiling can be applied to identify the cellular origin such as body fluids or organ tissues. The presence of male cell material can be readily assessed by the incorporation of Y-chromosomal markers in quantitation or STR profiling systems. However, no forensic marker exists to positively identify female cell material; merely the presence of female DNA is deduced from the absence of a Y peak, or unbalanced X-Y signals at the Amelogenin locus or unbalanced response of the total and Y-specific quantifier. The presence of two X-chromosomes in female cells invokes dosage compensation, which is achieved through inactivation of one of the X-chromosomes in females. Since this process involves specific RNA molecules, identification of female cellular material may be possible through RNA profiling. Additionally, male material may be identified through RNAs expressed from the Y-chromosome. RNAs preferentially expressed in either sex were assessed for their potential to act as sex markers in forensic RNA assays. To confirm sex-specificity, body fluids and organ tissues of multiple donors of either sex were tested. Additionally, sensitivity of the markers and the suitability of positively identifying male-female mixtures were assessed and degraded samples were used to assess performance of the markers in forensic settings. The addition of sex-specific markers is of added informative value in any RNA profiling system and both markers were incorporated into existing RNA assays that either target body fluids or organs. These are the first forensic assays that enable positive identification of female cellular material.
To read this article in full you will need to make a payment
Purchase one-time access:Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
One-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:Subscribe to Forensic Science International: Genetics
Already a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
- Forensic application of a rapid and quantitative DNA sex test by amplification of the X-Y homologous gene amelogenin.Int. J. Legal Med. 1994; 106: 190-193
- What do the X and Y chromosomes tell us about sex and gender in forensic case analysis?.J. Forensic Legal Med. 2007; 14: 27-30
- Structural variation on the short arm of the human Y chromosome: recurrent multigene deletions encompassing Amelogenin Y.Hum. Mol. Genet. 2007; 16: 307-316
- A rare mutation in the primer binding region of the amelogenin gene can interfere with gender identification.J. Mol. Diagn. 2016; 6: 401-405
- Application of Y-chromosomal STR haplotypes to forensic genetics.Croatian Med. J. 2001; 43: 292-297
- Developmental validation of the GlobalFiler® Express PCR Amplification Kit: a 6-dye multiplex assay for the direct amplification of reference samples.Forensic Sci. Int. Genet. 2015; 19: 148-155
- Developmental validation of the PowerPlex® fusion 6C system.Forensic Sci. Int. Genet. 2016; 21: 134-144
- A multiplex (m) RNA-profiling system for the forensic identification of body fluids and contact traces.Forensic Sci. Int.: Genet. 2012; 6: 565-577
- A collaborative European exercise on mRNA-based body fluid/skin typing and interpretation of DNA and RNA results.Forensic Sci. Int.: Genet. 2014; 10: 40-48
- mRNA profiling for body fluid identification by reverse transcription endpoint PCR and realtime PCR.Forensic Sci Int. Genet. 2009; 3: 80-88
- Highly specific mRNA biomarkers for the identification of vaginal secretions in sexual assault investigations.Sci. Justice. 2013; 53: 14-22
- Multiplex mRNA profiling for the identification of body fluids.Forensic Sci. Int. 2005; 152: 1-12
- mRNA profiling using a minimum of five mRNA markers per body fluid and a novel scoring method for body fluid identification.Int. J. Legal Med. 2013; 127: 707-721
- Molecular approaches for forensic cell type identification: on mRNA, miRNA, DNA methylation and microbial markers.Forensic Sci. Int.: Genet. 2015; 18: 21-32
- Advancing forensic RNA typing: on non-target secretions a nasal mucosa marker, a differential co-extraction protocol and the sensitivity of DNA and RNA profiling.Forensic Sci. Int.: Genet. 2016; 20: 119-129
- Development of a mRNA profiling multiplex for the inference of organ tissues.Int. J. Legal Med. 2013; 127: 891-900
- DNA and RNA profiling of excavated human remains with varying postmortem intervals.Int. J. Legal Med. 2016; 130: 1471-1480
- Robust and tissue-independent gender-specific transcript biomarkers.Biomarkers. 2013; 18: 436-445
- The evolution of sex-biased genes and sex-biased gene expression.Nat. Rev. Genet. 2007; 8: 689-698
- Long non-coding RNAs: a new frontier in the study of human diseases.Cancer Lett. 2013; 339: 159-166
- Long, abundantly expressed non-coding transcripts are altered in cancer.Hum. Mol. Genet. 2008; 17: 642-655
- Gender-specific gene expression in post-mortem human brain: localization to sex chromosomes.Neuropsychopharmacology. 2004; 29: 373
- Human and murine kidneys show gender-and species-specific gene expression differences in response to injury.PLoS One. 2009; 4: e4802
- The GTEx consortium, F.A., wright, R., lappalainen, M., calvo, G., getz, E.T., dermitzakis, K.G., ardlie, R. guigó, the human transcriptome across tissues and individuals.Science. 2015; 348: 660-665
- Xist RNA and the mechanism of X chromosome inactivation.Annu. Rev. Genet. 2002; 36: 233-278
- Xist and the order of silencing.EMBO Rep. 2007; 8: 34-39
- Long nonoding RNAs in the X-inactivation center.Chromosome Res. 2013; 21: 601-614
- No-nonsense functions for long noncoding RNAs.Cell. 2011; 145: 178-181
- Tsix, a gene antisense to Xist at the X-inactivation centre.Nat. Genet. 1999; 21: 400-404
- Requirement for Xist in X chromosome inactivation.Nature. 1996; 379: 131-137
- Cis-and trans-regulation in X inactivation.Chromosoma. 2016; 125: 41-50
- A self-enhanced transport mechanism through long noncoding RNAs for X chromosome inactivation.Sci. Rep. 2016; 6
- Genes that escape from X inactivation.Hum. Genet. 2011; 130: 237-245
- Silencing of the mammalian X chromosome.Annu. Rev. Genomics Hum. Genet. 2005; 6: 69-92
- Expression and function of a large non-coding RNA gene XIST in human cancer.World J. Surg. 2011; 35: 1751-1756
- The male-specific region of the human Y chromosome is a mosaic of discrete sequence classes.Nature. 2003; 423: 825-837
- Differential expression of biomarkers in men and women.Semin. Oncol. 2009; 36: 553-565
- Ensembl 2014.Nucleic Acids Res. 2014; 42: D749-D755
- PrimerBLAST: a tool to design target-specific primers for polymerase chain reaction.BMC Bioinf. 2012; 13: 134
- Implementation of RNA profiling in forensic casework.Forensic Sci. Int.: Genet. 2013; 7: 159-166
- RNA cell typing and DNA profiling of mixed samples: can cell types and donors be associated?.Sci. Justice. 2013; 53: 261-269
- Over-expression of XIST, the master gene for X chromosome inactivation, in females with major affective disorders.EBioMedicine. 2015; 2: 909-918
- Seven ring (X) chromosomes lacking the XIST locus, six with an unexpectedly mild phenotype.Hum. Genet. 2000; 106: 93-100
- A promoter mutation in the XIST gene in two unrelated families with skewed X-chromosome inactivation.Nat. Genet. 1997; 17: 353-356
Published online: October 26, 2016
Accepted: October 24, 2016
Received in revised form: October 17, 2016
Received: September 9, 2016
© 2016 Elsevier Ireland Ltd. All rights reserved.