Epithelial DLD-1 Cells with Disrupted E-cadherin Gene Retain the Ability to Form Cell Junctions and Apico-basal Polarity

• DOI: 10.1247/csf.15002
• Publication Date: 2015-08-17T18:02:34Z
• Authors: Miwako Fujiwara; Kihito Fujimura; Shuichi Obata; Ryo Yanagibashi; Tetsushi Sakuma; Takashi Yamamoto; Shintaro T. Suzuki
• Keywords: Polarity (international relations); Basal (medicine); Cell polarity

Gene editing methods were applied to the study of E-cadherin function in epithelial cells. The gene DLD-1 cells was ablated using TALEN. resultant showed round fibroblast-like morphology and had almost no Ca2+-dependent cell aggregation activity. re-expression knockout restored strong cell-cell adhesion activity, indicating that retained ability support cadherin function. partial localization desmoplakin ZO-1 at intercellular contact sites. transfectants expressing mutant lacking cytoplasmic domain clear sites, although only weak Electron microscopy revealed formation junctions apico-basal polarity these A portion occasionally formed an epithelial-like structure after prolonged culture. When treated with blebbistatin, enhanced. However, incomplete contained defects. Double-knockout MDCK for cadherin-6 genes similar results, suggesting above properties general. present results could be without E-cadherin, but construction mature epithelia requires E-cadherin.