Figure 2: Generation of mice carrying an HA-tag and a conditional allele for DOR.

A combination of TALEN mRNA (20 ng/μl, 1–2 pl/oocyte) and linearized DOR donor plasmid (20 ng/μl, 1–2 pl/oocyte) were injected into fertilized oocytes. After injection, the surviving zygotes were transplanted into foster mothers and postnatal mice were obtained and genotyped. (A) The picture of nine mice which were born after microinjection (No. 1–9). (B) PCR genotyping of the nine mice with F1, R1, F2, R2 primers. P, positive control (EGFP positive mESCs clone: C4). (C) Fluorescence image of the abdomen of the DOR^wt/wt (left top) and HA-DOR^floxETPP/wt (ETPP, EGFP-T2A-puro-PKG) mouse (No. 9, left bottom) and the F5 progeny of No. 9 mouse (right). (D) Nucleotide sequencing demonstrated HA insertion (red rectangles) in the correct location in the HA-DOR^floxETPP/wt mouse (No. 9). (E) PCR genotyping of the F1 progeny (No. 10–14) of mouse No. 9. (F) PCR analysis of LoxP and HA insertion with F3, R3 primers in DOR^wt/wt and No. 9 mouse.