The efficiency of gene knockin based on homology-directed repair (HDR) can be low, particularly in stem and primary cells. In addition, targeting a specific position in the genome can generally result in cells with untargeted insertion of the donor vector at non-specific locations or in carrying a cytoplasmic residue of the donor vector. Thus, we developed an improved method to select knocked-in cells efficiently based on donor vectors carrying double thymidine kinase of herpes simplex virus (HSV-tk) cassettes.
We have published this article in Cell Reports Methods (Nakade et al., Cell Reports Methods 2023).