An efficient and cost-effective method for disrupting genes in RAW264.7 macrophages using CRISPR-Cas9 v1

CRISPR-Cas9 genome editing is widely used to modify cultured cell lines. However, its implementation is still challenging due to the complex and often costly multi-step process associated with this technique and the variability in genome editing efficiency across cell types, which often limits utility. This protocol describes pCRISPR-EASY, a plasmid for simplified cloning of single guide RNAs and its simultaneous introduction with CRISPR-Cas9 into cultured cells using a non-viral delivery system. We outline a step-by-step protocol for genome editing in RAW264.7 macrophages, a mouse macrophage cell line that can easily be adapted for use in other systems.