Protocol for casein kinase 1γ3 CSNK1G3 gene knockout and recombinant gene expression in cultured HeLa cells

Casein kinase 1γ is a subfamily of the casein kinase 1 family. Here, we present a protocol for gene knockout of CSNK1G3 in human cells and cloning and/or expression of CSNK1G1–3 cDNAs using a retroviral vector system. We first describe the genome editing procedures, including sgRNA design, introduction into HeLa cells, and verification of genome editing. Next, we describe the procedures for cloning human CSNK1G cDNAs, introduction into HeLa cells, and expression verification by western blot analysis. For complete information on the generation and use of this protocol, please refer to Goto et al.(1)