TY - JOUR TI - A CRISPR screen in intestinal epithelial cells identifies novel factors for polarity and apical transport AU - Klee, Katharina MC AU - Hess, Michael W AU - Lohmüller, Michael AU - Herzog, Sebastian AU - Pfaller, Kristian AU - Müller, Thomas AU - Vogel, Georg F AU - Huber, Lukas A A2 - Bagnat, Michel A2 - Pfeffer, Suzanne R A2 - Martin-Belmonte, Fernando A2 - Bryant, David M VL - 12 PY - 2023 DA - 2023/01/20 SP - e80135 C1 - eLife 2023;12:e80135 DO - 10.7554/eLife.80135 UR - https://doi.org/10.7554/eLife.80135 AB - 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. KW - CRISPR KW - loss-of-function screen KW - epithelial polarity KW - membrane transport KW - electron microscopy KW - microvillus inclusion disease JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -