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Truong, D.J.J. ; Phlairaharn, T. ; Eßwein, B. ; Gruber, C. ; Tümen, D. ; Baligács, E. ; Armbrust, N. ; Vaccaro, F.L. ; Lederer, E.-M. ; Beck, E.M. ; Geilenkeuser, J. ; Göppert, S. ; Krumwiede, L. ; Grätz, C. ; Raffl, G. ; Schwarz, D. ; Zirngibl, M. ; Živanić, M. ; Beyer, M. ; Körner, J.D. ; Santl, T. ; Evsyukov, V.* ; Strauß, T.* ; Schwarz, S.C.* ; Höglinger, G.U.* ; Heutink, P.* ; Doll, S. ; Conrad, M. ; Giesert, F. ; Wurst, W. ; Westmeyer, G.G.

Non-invasive and high-throughput interrogation of exon-specific isoform expression.

Nat. Cell Biol. 23, 652-663 (2021)
Publ. Version/Full Text DOI PMC
Open Access Gold (Paid Option)
Creative Commons Lizenzvertrag
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.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Messenger-rna; Frontotemporal Dementia; Mapt Expression; Tau Exon-10; Activation; Mutations; Design; Gene; Parkinsonism; Inhibition
ISSN (print) / ISBN 1465-7392
e-ISSN 1476-4679
Journal Nature Cell Biology
Quellenangaben Volume: 23, Issue: 6, Pages: 652-663 Article Number: , Supplement: ,
Publisher Nature Publishing Group
Publishing Place Heidelberger Platz 3, Berlin, 14197, Germany
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Insitute of Synthetic Biomedicine (ISBM)
Institute of Developmental Genetics (IDG)
Institute of Metabolism and Cell Death (MCD)
Grants German Federal Ministry of Education and Research (BMBF) through EpiPD
TUM International Graduate School of Science and Engineering (IGSSE, project BIOMAG)
Bavarian Research Network for Molecular Biosystems (BioSysNet)
German Federal Ministry of Education and Research (BMBF) through HIT-Tau
AMPro project 'Aging and Metabolic Programming' of the Helmholtz Association
NOMIS foundation
Deutsche Forschungsgemeinschaft (DFG)
VolkswagenStiftung/Lower Saxony Ministry for Science (Etiology and Therapy of Synucleinopathies and Tauopathies)
Petermax-Muller Foundation
DFG
German Federal Ministry of Education and Research (BMBF) VIP+ program NEUROPROTEKT
Ministry of Science and Higher Education of The Russian Federation
Else Kroner-Fresenius-Stiftung
European Research Council (ERC) under the European Union
European Research Council