Highly scalable arrayed CRISPR mediated gene silencing in primary lung small airway epithelial cells

Abstract

Small airway epithelial cells (SAECs) play a central role in the pathogenesis of lung diseases and are now becoming a crucial cellular model for target identification and validation in drug discovery. However, primary cell lines such as SAECs are often difficult to transfect using traditional lipofection methods; therefore, gene editing using CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 is often carried out through ribonucleoprotein (RNP) electroporation. Here we have established a robust, scalable, and automated arrayed CRISPR nuclease (CRISPRn) screening workflow for SAECs which can be combined with a myriad of disease-specific endpoint assays.

Keywords

• CRISPR
• High throughput screening
• Arrayed screening
• Primary lung small airway epithelial cells
• CRISPR/Cas9
• SAECs