20.500.12556/DiRROS-15565 New approach for detection of normal alternative splicing events and aberrant spliceogenic transcripts with long-range PCR and deep RNA sequencing RNA sequencing is a promising technique for detecting normal and aberrant RNA isoforms. Here, we present a new single-gene, straightforward 1-day hands-on protocol for detection of splicing alterations with deep RNA sequencing from blood. We have validated our method%s accuracy by detecting previously published normal splicing isoforms of STK11 gene. Additionally, the same technique was used to provide the first comprehensive catalogue of naturally occurring alternative splicing events of the NBN gene in blood. Furthermore, we demonstrate that our approach can be used for detection of splicing impairment caused by genetic variants. Therefore, we were able to reclassify three variants of uncertain significance: NBN:c.584G>A, STK11:c.863-5_863-3delCTC and STK11:c.615G>A. Due to the simplicity of our approach, it can be incorporated into any molecular diagnostics laboratory for determination of variant%s impact on splicing. RNA sequencing DNA variant splicing sekvencioniranje RNA variante DNA spajanje true false true MDPI Angleški jezik Slovenski jezik by Authors Neznano 2022-09-21 03:35:12 2022-09-21 10:06:04 2022-09-30 15:14:39 0000-00-00 00:00:00 2021 0 0 Nasl. z nasl. zaslona; Opis vira z dne 31. 8. 2021; str. 706-1-706-12 no. 8 Vol. 10 2021 0000-00-00 Zaloznikova Objavljeno apc 0000-00-00 0000-00-00 2021-07-23 577 2079-7737 10.3390/biology10080706 74741763 https://www.mdpi.com/2079-7737/10/8/706 1 de042a2a-393c-11ed-bea5-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=20589 RAZ_Setrajcic_Dragos_Vita_2021.pdf RAZ_Setrajcic_Dragos_Vita_2021.pdf 1 BB715BB784D4A99A758173FC200A5B03 5d7638a0178f6e0c54f7a6522effa3e3f64fb39950ee9be41af0a6354b72d1ae a7f01a65-3973-11ed-bea5-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=20593 Onkološki inštitut Ljubljana 0 0 0