Developing Cas9 expressing MDBK cell lines for GeCKO screening against bovine diseases

Abstract

Cas9 expressing MDBK (Madin-Darby bovine kidney) cell lines were produced for a GeCKO (genome scale CRISPR/Cas9 gene knockout) screening against Bovine diseases. The lines were transduced with lentiviral vectors containing Cas9 and quantified for RNA expression and protein levels. sgRNAs were designed and assembled into lentiviral vectors. Transduction of Cas9 expressing MDBK cell lines with sgRNA containing lentivirus resulted in CRISPR/Cas9 gene editing. Editing efficiencies of selected lines were compared and correlated with their Cas9 expression profiles.