Figure 2.

TALEN-generated 750-bp and 4.2-kb deletions within the zebrafish apoea locus. (A) Schematic diagram of the apoea locus demarcating the positions of all four TALEN binding sites and the distance separating them. (B) Segmental deletions of 750 bp and 4.2 kb are 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 component. PCR products corresponding to the desired 750-bp and 4.2-kb segmental deletion products (∼517 and ∼402 bp, respectively) are detected only in embryos injected with both TALENs. Estimated deletion rates are indicated below the positive lanes. Segmental inversions are detected in embryos treated with two TALENs for both the 750-bp and 4.2-kb segments. The calculated inversion rates are lower than the corresponding segmental deletions. (C) Junction sequences for the 4.2-kb segmental deletions at the apoea locus. The TALEN-1 and TALEN-4 recognition sequences are indicated above the junction sequences, where the half-sites recognized by each TALEN are differentially colored. The recovered junction sequences contain variable length deletions as well as evidence of insertions between some of the target sites, where the latter are indicated in plain text.