Extended Data Fig. 8. ssDNA as donor template improves cell viability, but reduces KI efficiency.

a, KI CD4⁺ T cell cultures from Fig. 4b were lysed and a PCR specific for the eGFP integration into the SAMHD1 locus was performed. Untreated WT cells served as reference. A PCR specific for the CD46 locus was used as loading control (lower panel). b, GFP expression following KI (see also schematic in Fig. 4a). Density plots of flow cytometry analysis of viability (FSC/SSC, upper part) and GFP expression of resting CD4⁺ T cells two weeks after nucleofection. Cells were either left untreated (WT) or nucleofected with the indicated SAMHD1-gRNA2, dsDNA, ssDNA sense or ssDNA antisense templates alone or in combination. One µg of DNA template was used for each condition. One representative experiment is shown (n = 2).