Figure 5. Homologous recombination mediated gene targeting in hLR5 cells.
A: Schematic representation of the human HPRT locus and the targeting construct. The PCR primers used to detect the wild-type locus and the targeting construct (P1, P2, P3) are indicated.
B: PCR detection of 6 independent clones in which the HPRT locus was successfully targeted (KO1-6) and one clone with random integration of the targeting construct (RI). Wild-type cells (WT) were used as control. Upper panel, presence of the wild-type allele. Lower panel, detection of the targeting construct.
C: Confirmation of functional knockout of the HPRT gene in targeted hLR5 cells. WT: Wild-type, KO1 and KO2: knockout clones; RI: Clone with random integration, 6-TG: 6-thioguanine, HAT: HAT supplement