Fig. 4.

Generation of floxed mice at the Gpcpd1 locus. (A) Schematic illustration to generate a conditional allele at the Gpcpd1 locus. A 55-bp exon was targeted by two gRNA, which were designed to cut both ends of the exon. A single ssODN was designed to carry the two loxP sequences. Bold arrows indicate primers for PCR. Thin arrows indicate the recognition sites of restriction enzymes for the RFLP analysis. (B) Sequencing analysis of subcloned PCR1 products from three pups that were positive for all analyses. The intended floxed allele is shown at the top (Floxed) with loxP sequences (enclosed in black boxes). The wild-type allele is shown at the bottom. Dots indicate the same bases as the floxed sequence.