Highlights
- •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.
Abstract
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.
Keywords
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Article info
Publication history
Published online: October 26, 2016
Accepted:
October 24,
2016
Received in revised form:
October 17,
2016
Received:
September 9,
2016
Identification
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© 2016 Elsevier Ireland Ltd. All rights reserved.
