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Here we aimed to identify and validate mRNA candidates for a robust sub-differentiation of forensically and traumatologically relevant brain regions. To this purpose, massively parallel sequencing of whole transcriptomes in sample material taken from four different areas of the cerebral cortex (frontal, temporal, parietal, occipital lobe) was performed, followed by bioinformatical data analysis, classification, and biostatistical candidate selection. Candidates were evaluated by Multiplex-RT-PCR and capillary electrophoresis. Only a weak relative upregulation and solely for candidates expressed in the parietal lobe was observed. Two candidates with upregulation in the cerebellar region (PVALB and CDR2L) were chosen for further investigation; however, PVALB could not reliably and repeatedly be detected in any lobe whereas CDR2L was detectable in all lobes. Consequently, we suggest that differences in mRNA expression between four regions of the cerebral cortex are too small and less pronounced to be useful for and applicable in forensic RNA analysis. We conclude that sub-differentiation of these brain regions via RNA expression analysis is generally not feasible within a forensic scope.<\/jats:p>","DOI":"10.1007\/s12024-024-00787-7","type":"journal-article","created":{"date-parts":[[2024,1,31]],"date-time":"2024-01-31T13:02:41Z","timestamp":1706706161000},"page":"1276-1281","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Forensically relevant anatomical brain regions cannot be sub-differentiated by RNA expression analysis"],"prefix":"10.1007","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7800-3509","authenticated-orcid":false,"given":"Jan","family":"Euteneuer","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0700-1941","authenticated-orcid":false,"given":"Lucas","family":"Moitinho-Silva","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9811-8482","authenticated-orcid":false,"given":"Cornelius","family":"Courts","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,1,31]]},"reference":[{"key":"787_CR1","doi-asserted-by":"publisher","DOI":"10.1002\/wfs2.1407","author":"C Lynch","year":"2021","unstructured":"Lynch C, Fleming R. 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Development of a mRNA profiling multiplex for the inference of organ tissues. Int J Legal Med. 2013;127:891\u2013900. https:\/\/doi.org\/10.1007\/s00414-013-0895-7.","journal-title":"Int J Legal Med"},{"key":"787_CR4","doi-asserted-by":"publisher","first-page":"365","DOI":"10.1007\/s12024-015-9695-3","volume":"11","author":"M Grabm\u00fcller","year":"2015","unstructured":"Grabm\u00fcller M, Schyma C, Euteneuer J, Madea B, Courts C. Simultaneous analysis of nuclear and mitochondrial DNA, mRNA and miRNA from backspatter from inside parts of firearms generated by shots at \u201ctriple contrast\u201d doped ballistic models. Forensic Sci Med Pathol. 2015;11:365\u201375. https:\/\/doi.org\/10.1007\/s12024-015-9695-3.","journal-title":"Forensic Sci Med Pathol"},{"key":"787_CR5","doi-asserted-by":"publisher","DOI":"10.1016\/j.forsciint.2020.110632","volume":"318","author":"C Schyma","year":"2021","unstructured":"Schyma C, Madea B, M\u00fcller R, Zieger M, Utz S, Grabm\u00fcller M. 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The study design and experimental procedures were approved by the ethics committees of the University Hospital of Bonn and the University Medical Center of Schleswig-Holstein.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"Preliminary results from this study were presented at the 29th Congress of the International Society for Forensic Genetics, Washington, DC, USA, August 29th to September 2nd, 2022 (P209). Concomitantly, a non-peer-reviewed \u201cextended abstract\u201d titled \u201cTowards localizing head shots \u2013 Forensic sub-differentiation of anatomical brain regions by differential RNA expression\u201d was published in the Forensic Science International: Genetics Supplement Series (), the corresponding publication venue for the proceedings of scientific symposia.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Previous or partial publication of contents"}}]}}