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. 2013 Apr 30;21(6):1232–1241. doi: 10.1038/mt.2013.59

Figure 2.

Figure 2

B2M targeting and transgene removal. (a) The structures of the AAV3-B2M-ETKNpA– and AAV3-EHyTKpA–targeting vectors, and the human B2M locus are shown before targeting, and after two rounds of targeting and transgene removal by Cre. The locations of Southern blot probes and enzymes (X, Xba I) are indicated. (b) Diagram of the wild-type, targeted, and transgene-deleted alleles of the B2M gene with the locations of PCR primers, and the results obtained from an analysis of 45 Cre-treated clones. The gel below shows a representative PCR analysis of Cre-transduced clones with the different primer combinations, and the resulting allele designations are shown. DNA from the parental H1 cells (+/+), and clones targeted in one (+/TKNeo) or both (HyTK/TKNeo) B2M alleles were used as controls. (c) Southern blot analysis of Xba I–digested genomic DNAs of parental H1 cells (+/+), one clone targeted at one B2M allele (+/TKNeo), one clone targeted at both B2M alleles (HyTK/TKNeo), and four clones obtained after removal of the transgenes with Cre (loxP/loxP c1–4), probed with B2M- or TK-specific probes. Positions of the different possible B2M fragments are shown at the left.