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Article

Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing

Department of Forensic Genetics, Institute of Legal Medicine, Ludwig Maximilian University of Munich, Nußbaumstraße 26, 80336 Munich, Germany
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Author to whom correspondence should be addressed.
Academic Editor: Yutaka Suzuki
Genes 2021, 12(9), 1362; https://doi.org/10.3390/genes12091362
Received: 28 July 2021 / Revised: 24 August 2021 / Accepted: 27 August 2021 / Published: 30 August 2021
(This article belongs to the Special Issue Advances in Forensic Genetics)
Single-cell sequencing is a fast developing and very promising field; however, it is not commonly used in forensics. The main motivation behind introducing this technology into forensics is to improve mixture deconvolution, especially when a trace consists of the same cell type. Successful studies demonstrate the ability to analyze a mixture by separating single cells and obtaining CE-based STR profiles. This indicates a potential use of the method in other forensic investigations, like forensic DNA phenotyping, in which using mixed traces is not fully recommended. For this study, we collected single-source autopsy blood from which the white cells were first stained and later separated with the DEPArray™ N×T System. Groups of 20, 10, and 5 cells, as well as 20 single cells, were collected and submitted for DNA extraction. Libraries were prepared using the Ion AmpliSeq™ PhenoTrivium Panel, which includes both phenotype (HIrisPlex-S: eye, hair, and skin color) and ancestry-associated SNP-markers. Prior to sequencing, half of the single-cell-based libraries were additionally amplified and purified in order to improve the library concentrations. Ancestry and phenotype analysis resulted in nearly full consensus profiles resulting in correct predictions not only for the cells groups but also for the ten re-amplified single-cell libraries. Our results suggest that sequencing of single cells can be a promising tool used to deconvolute mixed traces submitted for forensic DNA phenotyping. View Full-Text
Keywords: forensic DNA phenotyping; FDP; HIrisPlex-S; DEPArray; ancestry prediction; phenotype prediction; massively parallel sequencing; next-generation sequencing; single-cell genomics; single-cell sequencing; mixture deconvolution; low template DNA; ltDNA forensic DNA phenotyping; FDP; HIrisPlex-S; DEPArray; ancestry prediction; phenotype prediction; massively parallel sequencing; next-generation sequencing; single-cell genomics; single-cell sequencing; mixture deconvolution; low template DNA; ltDNA
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MDPI and ACS Style

Diepenbroek, M.; Bayer, B.; Anslinger, K. Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing. Genes 2021, 12, 1362. https://doi.org/10.3390/genes12091362

AMA Style

Diepenbroek M, Bayer B, Anslinger K. Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing. Genes. 2021; 12(9):1362. https://doi.org/10.3390/genes12091362

Chicago/Turabian Style

Diepenbroek, Marta, Birgit Bayer, and Katja Anslinger. 2021. "Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing" Genes 12, no. 9: 1362. https://doi.org/10.3390/genes12091362

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