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Truong, Dong-Jiunn Jeffery; Phlairaharn, Teeradon; Eßwein, Bianca; Gruber, Christoph; Tümen, Deniz; Baligács, Enikő; Armbrust, Niklas; Vaccaro, Francesco Leandro; Lederer, Eva-Maria; Beck, Eva Magdalena; Geilenkeuser, Julian; Göppert, Simone; Krumwiede, Luisa; Grätz, Christian; Raffl, Gerald; Schwarz, Dominic; Zirngibl, Martin; Živanić, Milica; Beyer, Maren; Körner, Johann Dietmar; Santl, Tobias; Evsyukov, Valentin; Strauß, Tabea; Schwarz, Sigrid C.; [...]

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

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  • Media type: E-Article
  • Title: Non-invasive and high-throughput interrogation of exon-specific isoform expression
  • Contributor: Truong, Dong-Jiunn Jeffery; Phlairaharn, Teeradon; Eßwein, Bianca; Gruber, Christoph; Tümen, Deniz; Baligács, Enikő; Armbrust, Niklas; Vaccaro, Francesco Leandro; Lederer, Eva-Maria; Beck, Eva Magdalena; Geilenkeuser, Julian; Göppert, Simone; Krumwiede, Luisa; Grätz, Christian; Raffl, Gerald; Schwarz, Dominic; Zirngibl, Martin; Živanić, Milica; Beyer, Maren; Körner, Johann Dietmar; Santl, Tobias; Evsyukov, Valentin; Strauß, Tabea; Schwarz, Sigrid C.; [...]
  • imprint: Springer Science and Business Media LLC, 2021
  • Published in: Nature Cell Biology
  • Language: English
  • DOI: 10.1038/s41556-021-00678-x
  • ISSN: 1465-7392; 1476-4679
  • Origination:
  • Footnote:
  • Description: <jats:title>Abstract</jats:title><jats:p>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 (<jats:italic>MAPT</jats:italic>) 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 <jats:italic>FOXP1</jats:italic>, 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.</jats:p>