Epithelial miR-141 regulates IL-13-induced airway mucus production

We performed RNA sequencing of gene expression in primary human bronchial epithelial cells that have undergone CRISPR/Cas9-based targeting of MIR141. The goal was to identify the role of miR-141 in goblet cell mucus production. CRISPR-targeted cells were differentiated at air-liquid-interface and stimulated with IL-13 to induce goblet cell hyperplasia. An electroporation-based dual guide RNA (gRNA; cRNA:tracrRNA) CRISPR protocol enabled MIR141 gene repression in primary human bronchial epithelial cells (HBECs) grown in monolayer cultures. Subsequent transfer to air-liquid-interface generated a fully differentiated airway epithelium. We harvested IL-13-stimulated HBECs that received either MIR141 gene-targeting gRNA (n=4) or non-targeting (NT) gRNA control (n=4) and untreated HBECs that received either MIR141 gene-targeting gRNA (n=3) or NT gRNA control (n=3).