دورية أكاديمية
A CRISPR screen in intestinal epithelial cells identifies novel factors for polarity and apical transport
| العنوان: | A CRISPR screen in intestinal epithelial cells identifies novel factors for polarity and apical transport |
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| المؤلفون: | Katharina MC Klee, Michael W Hess, Michael Lohmüller, Sebastian Herzog, Kristian Pfaller, Thomas Müller, Georg F Vogel, Lukas A Huber |
| المصدر: | eLife, Vol 12 (2023) |
| بيانات النشر: | eLife Sciences Publications Ltd, 2023. |
| سنة النشر: | 2023 |
| المجموعة: | LCC:Medicine LCC:Science LCC:Biology (General) |
| مصطلحات موضوعية: | CRISPR, loss-of-function screen, epithelial polarity, membrane transport, electron microscopy, microvillus inclusion disease, Medicine, Science, Biology (General), QH301-705.5 |
| الوصف: | Epithelial polarization and polarized cargo transport are highly coordinated and interdependent processes. In our search for novel regulators of epithelial polarization and protein secretion, we used a genome-wide CRISPR/Cas9 screen and combined it with an assay based on fluorescence-activated cell sorting (FACS) to measure the secretion of the apical brush-border hydrolase dipeptidyl peptidase 4 (DPP4). In this way, we performed the first CRISPR screen to date in human polarized epithelial cells. Using high-resolution microscopy, we detected polarization defects and mislocalization of DPP4 to late endosomes/lysosomes after knockout of TM9SF4, anoctamin 8, and ARHGAP33, confirming the identification of novel factors for epithelial polarization and apical cargo secretion. Thus, we provide a powerful tool suitable for studying polarization and cargo secretion in epithelial cells. In addition, we provide a dataset that serves as a resource for the study of novel mechanisms for epithelial polarization and polarized transport and facilitates the investigation of novel congenital diseases associated with these processes. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2050-084X |
| Relation: | https://elifesciences.org/articles/80135; https://doaj.org/toc/2050-084X |
| DOI: | 10.7554/eLife.80135 |
| URL الوصول: | https://doaj.org/article/8df03ae96a5f4605be497b42d8888f95 |
| رقم الأكسشن: | edsdoj.8df03ae96a5f4605be497b42d8888f95 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 2050084X |
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| DOI: | 10.7554/eLife.80135 |