CRISPR/Cas9 editing of Primary Bovine Mammary Epithelial Cells

This project assessed four pbMEC lines recovered from pregnant cows. we describe an optimised method for preferentially selecting CRISPR-Cas9 edited cells conferring a knock out of DGAT1, using fluorescence activated cell sorting (FACS). Editing efficiency was determined by minION sequencing of a PCR amplicon of the edited region. The combination of these techniques facilitate the use of pbMECs as a model to investigate the effects of gene introgressions and genetic variation in lactating mammary tissue.