Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing
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 d...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-08-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/12/9/1362 |
| _version_ | 1797519143333789696 |
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| author | Marta Diepenbroek Birgit Bayer Katja Anslinger |
| author_facet | Marta Diepenbroek Birgit Bayer Katja Anslinger |
| author_sort | Marta Diepenbroek |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T07:38:54Z |
| format | Article |
| id | doaj.art-a273b7604bc64b61a919c67fbb8266c4 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-10T07:38:54Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-a273b7604bc64b61a919c67fbb8266c42023-11-22T13:13:54ZengMDPI AGGenes2073-44252021-08-01129136210.3390/genes12091362Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell SequencingMarta Diepenbroek0Birgit Bayer1Katja Anslinger2Department of Forensic Genetics, Institute of Legal Medicine, Ludwig Maximilian University of Munich, Nußbaumstraße 26, 80336 Munich, GermanyDepartment of Forensic Genetics, Institute of Legal Medicine, Ludwig Maximilian University of Munich, Nußbaumstraße 26, 80336 Munich, GermanyDepartment of Forensic Genetics, Institute of Legal Medicine, Ludwig Maximilian University of Munich, Nußbaumstraße 26, 80336 Munich, GermanySingle-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.https://www.mdpi.com/2073-4425/12/9/1362forensic DNA phenotypingFDPHIrisPlex-SDEPArrayancestry predictionphenotype prediction |
| spellingShingle | Marta Diepenbroek Birgit Bayer Katja Anslinger Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing Genes forensic DNA phenotyping FDP HIrisPlex-S DEPArray ancestry prediction phenotype prediction |
| title | Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing |
| title_full | Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing |
| title_fullStr | Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing |
| title_full_unstemmed | Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing |
| title_short | Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing |
| title_sort | pushing the boundaries forensic dna phenotyping challenged by single cell sequencing |
| topic | forensic DNA phenotyping FDP HIrisPlex-S DEPArray ancestry prediction phenotype prediction |
| url | https://www.mdpi.com/2073-4425/12/9/1362 |
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