Truong, Dong-Jiunn Jeffery and Phlairaharn, Teeradon and Esswein, Bianca and Gruber, Christoph and Tuemen, Deniz and Baligacs, Eniko and Armbrust, Niklas and Vaccaro, Francesco Leandro and Lederer, Eva-Maria and Beck, Eva Magdalena and Geilenkeuser, Julian and Goeppert, Simone and Krumwiede, Luisa and Graetz, Christian and Raffl, Gerald and Schwarz, Dominic and Zirngibl, Martin and Zivanic, Milica and Beyer, Maren and Koerner, Johann Dietmar and Santl, Tobias and Evsyukov, Valentin and Strauss, Tabea and Schwarz, Sigrid C. and Hoeglinger, Guenter U. and Heutink, Peter and Doll, Sebastian and Conrad, Marcus and Giesert, Florian and Wurst, Wolfgang and Westmeyer, Gil Gregor (2021) Non-invasive and high-throughput interrogation of exon-specific isoform expression. NATURE CELL BIOLOGY, 23 (6). 652-+. ISSN 1465-7392, 1476-4679
Full text not available from this repository. (Request a copy)Abstract
Truong et al. developed a cell-based reporter system, EXSISERS, that enables non-invasive quantification of the protein expression levels of exon-specific isoforms via intein-mediated protein splicing. Expression of exon-specific isoforms from alternatively spliced mRNA is a fundamental mechanism that substantially expands the proteome of a cell. However, conventional methods to assess alternative splicing are either consumptive and work-intensive or do not quantify isoform expression longitudinally at the protein level. Here, we therefore developed an exon-specific isoform expression reporter system (EXSISERS), which non-invasively reports the translation of exon-containing isoforms of endogenous genes by scarlessly excising reporter proteins from the nascent polypeptide chain through highly efficient, intein-mediated protein splicing. We applied EXSISERS to quantify the inclusion of the disease-associated exon 10 in microtubule-associated protein tau (MAPT) in patient-derived induced pluripotent stem cells and screened Cas13-based RNA-targeting effectors for isoform specificity. We also coupled cell survival to the inclusion of exon 18b of FOXP1, which is involved in maintaining pluripotency of embryonic stem cells, and confirmed that MBNL1 is a dominant factor for exon 18b exclusion. EXSISERS enables non-disruptive and multimodal monitoring of exon-specific isoform expression with high sensitivity and cellular resolution, and empowers high-throughput screening of exon-specific therapeutic interventions.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | MESSENGER-RNA; FRONTOTEMPORAL DEMENTIA; MAPT EXPRESSION; TAU EXON-10; ACTIVATION; MUTATIONS; DESIGN; GENE; PARKINSONISM; INHIBITION; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin I |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 21 Sep 2022 14:13 |
| Last Modified: | 21 Sep 2022 14:13 |
| URI: | https://pred.uni-regensburg.de/id/eprint/47873 |
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