The human TRAM1 locus expresses circular RNAs
Abstract Numerous indirect and in silico produced evidences suggest circular RNAs (circRNA) in mammals while thorough experimental proofs of their existence have rarely been reported. Biological studies of circRNA, however, should be based on experimentally verified circRNAs. Here, we describe the i...
| Main Authors: | , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2021-11-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-01548-0 |
| _version_ | 1798034927283863552 |
|---|---|
| author | Josephine Dubois Georg Sczakiel |
| author_facet | Josephine Dubois Georg Sczakiel |
| author_sort | Josephine Dubois |
| collection | DOAJ |
| description | Abstract Numerous indirect and in silico produced evidences suggest circular RNAs (circRNA) in mammals while thorough experimental proofs of their existence have rarely been reported. Biological studies of circRNA, however, should be based on experimentally verified circRNAs. Here, we describe the identification of two circRNAs originating from the gene locus of the translocation associated membrane protein 1 (TRAM1). Linear and potentially circular TRAM1-specific transcripts were identified in a transcriptome analysis of urine RNA of bladder cancer (BCa) patients versus healthy donors. Thus, we first focused on the topology of TRAM1-specific transcripts. We describe conclusive experimental evidence for the existence of TRAM1-specific circRNAs in the human BCa cell lines ECV-304 and RT-4. PCR-based methodology followed by cloning and sequencing strongly indicated the circular topology of two TRAM1 RNAs. Further, studies with exon fusion sequence-specific antisense oligonucleotides (asON) and RNase H as well as studies in the use of RNase R contribute to conclusive set of experiments supporting the circular topology of TRAM1 transcripts. On the biological side, TRAM1-specific circRNAs showed low expression levels and minor differences in BCa cell lines while linear TRAM1 transcripts displayed down-regulated expression in the higher cancer stage model ECV-304 versus more differentiated RT-4 cells. |
| first_indexed | 2024-04-11T20:51:10Z |
| format | Article |
| id | doaj.art-99c3be04329a4556ac01ef5622f903d7 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T20:51:10Z |
| publishDate | 2021-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-99c3be04329a4556ac01ef5622f903d72022-12-22T04:03:51ZengNature PortfolioScientific Reports2045-23222021-11-0111111510.1038/s41598-021-01548-0The human TRAM1 locus expresses circular RNAsJosephine Dubois0Georg Sczakiel1Institut für Molekulare Medizin, Universität zu Lübeck and UKSHInstitut für Molekulare Medizin, Universität zu Lübeck and UKSHAbstract Numerous indirect and in silico produced evidences suggest circular RNAs (circRNA) in mammals while thorough experimental proofs of their existence have rarely been reported. Biological studies of circRNA, however, should be based on experimentally verified circRNAs. Here, we describe the identification of two circRNAs originating from the gene locus of the translocation associated membrane protein 1 (TRAM1). Linear and potentially circular TRAM1-specific transcripts were identified in a transcriptome analysis of urine RNA of bladder cancer (BCa) patients versus healthy donors. Thus, we first focused on the topology of TRAM1-specific transcripts. We describe conclusive experimental evidence for the existence of TRAM1-specific circRNAs in the human BCa cell lines ECV-304 and RT-4. PCR-based methodology followed by cloning and sequencing strongly indicated the circular topology of two TRAM1 RNAs. Further, studies with exon fusion sequence-specific antisense oligonucleotides (asON) and RNase H as well as studies in the use of RNase R contribute to conclusive set of experiments supporting the circular topology of TRAM1 transcripts. On the biological side, TRAM1-specific circRNAs showed low expression levels and minor differences in BCa cell lines while linear TRAM1 transcripts displayed down-regulated expression in the higher cancer stage model ECV-304 versus more differentiated RT-4 cells.https://doi.org/10.1038/s41598-021-01548-0 |
| spellingShingle | Josephine Dubois Georg Sczakiel The human TRAM1 locus expresses circular RNAs Scientific Reports |
| title | The human TRAM1 locus expresses circular RNAs |
| title_full | The human TRAM1 locus expresses circular RNAs |
| title_fullStr | The human TRAM1 locus expresses circular RNAs |
| title_full_unstemmed | The human TRAM1 locus expresses circular RNAs |
| title_short | The human TRAM1 locus expresses circular RNAs |
| title_sort | human tram1 locus expresses circular rnas |
| url | https://doi.org/10.1038/s41598-021-01548-0 |
| work_keys_str_mv | AT josephinedubois thehumantram1locusexpressescircularrnas AT georgsczakiel thehumantram1locusexpressescircularrnas AT josephinedubois humantram1locusexpressescircularrnas AT georgsczakiel humantram1locusexpressescircularrnas |