TEQUILA-seq : a versatile and low-cost method for targeted long-read RNA sequencing
© 2023. Springer Nature Limited..
Long-read RNA sequencing (RNA-seq) is a powerful technology for transcriptome analysis, but the relatively low throughput of current long-read sequencing platforms limits transcript coverage. One strategy for overcoming this bottleneck is targeted long-read RNA-seq for preselected gene panels. We present TEQUILA-seq, a versatile, easy-to-implement, and low-cost method for targeted long-read RNA-seq utilizing isothermally linear-amplified capture probes. When performed on the Oxford nanopore platform with multiple gene panels of varying sizes, TEQUILA-seq consistently and substantially enriches transcript coverage while preserving transcript quantification. We profile full-length transcript isoforms of 468 actionable cancer genes across 40 representative breast cancer cell lines. We identify transcript isoforms enriched in specific subtypes and discover novel transcript isoforms in extensively studied cancer genes such as TP53. Among cancer genes, tumor suppressor genes (TSGs) are significantly enriched for aberrant transcript isoforms targeted for degradation via mRNA nonsense-mediated decay, revealing a common RNA-associated mechanism for TSG inactivation. TEQUILA-seq reduces the per-reaction cost of targeted capture by 2-3 orders of magnitude, as compared to a standard commercial solution. TEQUILA-seq can be broadly used for targeted sequencing of full-length transcripts in diverse biomedical research settings.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:14 |
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| Enthalten in: |
Nature communications - 14(2023), 1 vom: 08. Aug., Seite 4760 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wang, Feng [VerfasserIn] |
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| Links: |
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| Themen: |
63231-63-0 |
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| Anmerkungen: |
Date Completed 10.08.2023 Date Revised 12.02.2024 published: Electronic Citation Status MEDLINE |
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| doi: |
10.1038/s41467-023-40083-6 |
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| Förderinstitution / Projekttitel: |
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| 520 | |a Long-read RNA sequencing (RNA-seq) is a powerful technology for transcriptome analysis, but the relatively low throughput of current long-read sequencing platforms limits transcript coverage. One strategy for overcoming this bottleneck is targeted long-read RNA-seq for preselected gene panels. We present TEQUILA-seq, a versatile, easy-to-implement, and low-cost method for targeted long-read RNA-seq utilizing isothermally linear-amplified capture probes. When performed on the Oxford nanopore platform with multiple gene panels of varying sizes, TEQUILA-seq consistently and substantially enriches transcript coverage while preserving transcript quantification. We profile full-length transcript isoforms of 468 actionable cancer genes across 40 representative breast cancer cell lines. We identify transcript isoforms enriched in specific subtypes and discover novel transcript isoforms in extensively studied cancer genes such as TP53. Among cancer genes, tumor suppressor genes (TSGs) are significantly enriched for aberrant transcript isoforms targeted for degradation via mRNA nonsense-mediated decay, revealing a common RNA-associated mechanism for TSG inactivation. TEQUILA-seq reduces the per-reaction cost of targeted capture by 2-3 orders of magnitude, as compared to a standard commercial solution. TEQUILA-seq can be broadly used for targeted sequencing of full-length transcripts in diverse biomedical research settings | ||
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| 700 | 1 | |a Wang, Robert |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Beatrice |e verfasserin |4 aut | |
| 700 | 1 | |a Smith, Noah |e verfasserin |4 aut | |
| 700 | 1 | |a Notaro, Amber |e verfasserin |4 aut | |
| 700 | 1 | |a Gaerlan, Samantha |e verfasserin |4 aut | |
| 700 | 1 | |a Kutschera, Eric |e verfasserin |4 aut | |
| 700 | 1 | |a Kadash-Edmondson, Kathryn E |e verfasserin |4 aut | |
| 700 | 1 | |a Xing, Yi |e verfasserin |4 aut | |
| 700 | 1 | |a Lin, Lan |e verfasserin |4 aut | |
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