Figure 5.

TALEN-generated deletions of a segment of the globin LCR on chromosome 3 and the linc-birc6 (megamind) on chromosome 17. (A) Schematic diagram of the nprl3 locus demarcating the positions of the two TALEN binding sites flanking the core LCR regulatory element within this intron and the distance separating them. (B) Segmental deletion of the 300-bp element is detected in normal-appearing TALEN-treated embryos when two TALENs are coinjected. The presence or absence of each TALEN component (5′ and 3′ recognition unit) is indicated above each gel lane, as is the dose of each TALEN component. A PCR product corresponding to the desired segmental deletion product is detected only in embryos injected with both TALENs. Estimated deletion rates are indicated below the positive lanes. (C) Junction sequences for the segmental deletion of this element are shown below, where there are variable length deletions between the distal TALEN recognition sites. (D) Schematic diagram of the birc6 locus demarcating the positions of the two TALEN binding sites flanking the linc-birc6 (megamind) lincRNA gene and the distance separating them. (E) Segmental deletion of the 3.2-kb element is readily detected in normal-appearing TALEN-treated embryos when two TALENs are coinjected. The presence or absence of each TALEN component (5′ and 3′ recognition unit) is indicated above each gel lane, as is the dose of each TALEN component. A PCR product corresponding to the desired segmental deletion product is detected only in embryos injected with both TALENs. The estimated deletion rate is indicated below the positive lane. (F) Junction sequences for the 3.2-kb segmental deletions of the linc-birc6 (megamind). The TALEN-3 and TALEN-6 recognition sequences are indicated above the junction sequences, where the TALEN half-sites are differentially colored. In the recovered junction sequences, there are variable length deletions.