Fig. 2. Comparison of TALEN- and CRISPR/Cas9-mediated HDR at the yellow locus.

(A) CRISPR/Cas9- or TALEN-mediated yellow deletion. The CRISPR/Cas9 binding sequence is 5′-GGGTTTTGGACACTGGAACCG-3′ (underlined in panel A); PAM sequence is marked in red. One pair of TALEN binding sites is marked by boxes. The pairs of scissors indicate the Cas9 or TALEN cutting site at the yellow locus. The dashed red lines represent the deleted yellow genomic sequence (0.18 kb). (B) Molecular identification of the yellow deletions. The genomic DNAs of two heterozygous F₁ lines, 46-3 (CRISPR/Cas9-mediated mutagenesis) and 8-1 (TALEN-mediated mutagenesis) were used as examples. The pair of primers used for PCR is shown in panel A (opposing arrows). The appearance of a 0.46 kb PCR band indicates successful deletion. (C) Frequencies of CRISPR/Cas9- and TALEN-mediated HDR at the yellow locus. The deletion-yielding events in both F₀ and F₁ are scored based on the appearance of the 0.64 kb short PCR product shown in panel B. The legends for the rest of the elements/labels are the same as in Fig. 1.