TY - JOUR T1 - A CRISPR-screen in intestinal epithelial cells identifies novel factors for polarity and apical transport JF - bioRxiv DO - 10.1101/2022.05.16.492077 SP - 2022.05.16.492077 AU - Katharina MC Klee AU - Michael W Hess AU - Michael Lohmüller AU - Sebastian Herzog AU - Kristian Pfaller AU - Thomas Müller AU - Georg F Vogel AU - Lukas A Huber Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/05/17/2022.05.16.492077.abstract N2 - 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 knock-down 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.Competing Interest StatementThe authors have declared no competing interest.CaCo2caucasian colorectal adenocarcinoma 2CODEcongenital diarrhea and enteropathyDAMP3-(2,4-Dinitroanilino)-3’amino-N-methylpropyl-amineDPP4Dipeptidylpeptidase 4GOgene ontologyKDknock-downKEGGKyoto Encyclopedia of Genes and GenomesMDCKmadine darby canine kidneyMVImicrovillus inclusionMVIDmicrovillus inclusion diseaseTEERtransepithelial electric resistance ER -