Enhancement of microhomology-mediated genomic rearrangements by transient loss of mouse Bloom syndrome helicase

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Figure 2.
Figure 2.

Polymorphic marker analyses of chromosome 8 in Blmtet/tetAprtneo/+ ES cell clones selected by 2-FA for LOH at the Aprt locus. (A) (Right) Twenty-nine independent Dox(+)ISceIo(–) ES cell clones resistant to 2-FA (numbered as indicated at the top) were genotyped for 12 SNPs (SNP1 to SNP12; SNP4 is not shown because of the low specificity of the PCR reaction) covering chromosome 8 (Supplemental Table S1). Open squares indicate the heterozygous genotype (B6/129). Closed black and red squares indicate LOH with retention of the B6 and 129 alleles, respectively. (BA) Control 2-FA-unselected Dox(–)ISceIo(–) Blmtet/tetAprtneo/+ ES cells. (Left) Histogram representation of LOH ratio (total number of clones with LOH/total number of clones examined × 100 [%]) at each marker locus. (B) (Right) Sixty-four independent Dox(+)ISceIo(+) ES cell clones resistant to 2-FA (numbered as indicated at the top) were genotyped for SNP1 to SNP12. The 64 clones are divided into two groups: Group I, 55 clones carrying LOH with retention of the B6 allele (LOHB6) at the SNP12 locus; Group II, the other nine clones carrying no LOH or LOH129 at the SNP12 locus. (Left) Histogram representation of LOH ratio (%) at each marker locus of Group I clones. (C) Flow diagram of the SNP analysis performed in this study. The number of independent clones analyzed is shown in parentheses. (D) Detailed SNP analysis of the region between SNP11 and SNP12 in the clones shown in B. An additional five SNPs (u-50k, u-10k, u-5k, d-6k, and d-50k) were genotyped (Supplemental Table S1). (E) Ten independent Dox(–)ISceIo(+) ES cell clones resistant to 2-FA (numbered and categorized into Group Ic or IIc as indicated) were genotyped for SNP1 to SNP12. In the following experiments, clones indicated by asterisks at the bottom (D and E) carry DELs involving this region. Clone II-73 (dagger) carries a missense mutation at exon 1 of the 129-derived Aprt gene. Clone IIc-46 (double dagger) retains heterozygosity at the Aprt locus, and the mutation in the 129 allele is undefined.

This Article

  1. Published in Advance August 1, 2013, doi: 10.1101/gr.152744.112 Genome Res. 23: 1462-1473

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