New approach for detection of normal alternative splicing events and aberrant spliceogenic transcripts with long-range PCR and deep RNA sequencing
| Autor: | Vida Stegel, Srdjan Novaković, Mateja Krajc, Gašper Klančar, Ana Blatnik, Vita Setrajcic Dragos |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Gene isoform QH301-705.5 STK11 Computational biology DNA variant Biology Article General Biochemistry Genetics and Molecular Biology germline variant 03 medical and health sciences alternative splicing splicing 0302 clinical medicine variante DNA RNA Isoforms udc:577 Biology (General) Gene splicing variant General Immunology and Microbiology sekvencioniranje RNA spliceogenic variant Alternative splicing RNA RNA sequencing Molecular diagnostics spajanje 030104 developmental biology sekvencioniranje RNA variante DNA spajanje 030220 oncology & carcinogenesis RNA splicing alternative transcript RNA sequencing DNA variant splicing General Agricultural and Biological Sciences |
| Zdroj: | Biology, vol. 10, no. 8, pp. 706-1-706-12, 2021. Biology, Vol 10, Iss 706, p 706 (2021) Biology, str. 706-1-706-12, Vol. 10, no. 8, 2021 COBISS-ID: 523004441 Biology Volume 10 Issue 8 |
| ISSN: | 2079-7737 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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