Table 1.
Efficiency of targeted insertion of the HSV1tk-Venus fusion gene into the GAPDH locus with CRISPR/Cas9-mediated genome editing.
| Human iPSCs | No. of FACS-Sorted Venus-Positive Cells/Well | ||
|---|---|---|---|
| 1 | 10 | 100 | |
| (No. of Surviving Cells/Total No. of Sorted Cells) | |||
| 253G1 | 0/26 | 0/10 | 1*/100 |
| 1231A3 | 1*/26 | 0/10 | 1*/100 |
| 1210B2 | 0/26 | 0/10 | 1*/200 |
| 1210B2 (Cont.) | 6/24 | 2/20 | 20/200 |
Human iPSCs (253G1, 1231A3, and 1210B2) were subjected to transfection with the pU6-GAPDHgRNA4-Cas9 plasmid and the HR-GAPDH-2A-HSV1tk-2A-Venus donor plasmid. The indicated numbers of Venus-positive cells were FACS-sorted into individual wells of a 96-well plate. The numbers of surviving cells per the total number of FACS-sorted cells are shown. *, Venus negative cells; Cont., HR-GAPDH-2A-Venus donor plasmid was used as a control.