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. 2015 Sep 14;5:14057. doi: 10.1038/srep14057

Figure 2. The dosage effects and efficiency evaluation of DMY-TALENs.

Figure 2

(A) Microinjection of TALENs mRNA into medaka. (B) Detection of mutations in TALEN targeted medaka embryos. Primers DMY F, DMY F1 and DMY R were used to amply the DMY gene. Primers DMY F and DMY R bridge both EBE regions, while primers DMY F1 and DMY R link the spacer region and the downstream EBE. DMY F and DMY R generated a 396 bp PCR fragment. DMY F1 and DMY R generated a 167 bp PCR fragment. If primer DMY F and DMY R generated a PCR fragment, while primer DMY F1 and DMY R failed to do so, this suggested that the targeted gene is disrupted by the DMY-TALEN. (C) Electrophoretic detection of mutations in TALEN-injected medaka embryos. Line 1 to 16, TALENs injected embryos. 1,3,6,11,14 show mutated embryos. (D) Sequencing detection of mutations in TALEN-induced medaka embryos. -, deleted nucleotide; lowercase letter, added nucleotide. +, insertions; Δ, deletions. (E) Evaluation of embryonic toxicity of DMY-TALENs. (F) Evaluation of embryonic toxicity of DMY-nanos3UTR-TALENs. (G) The targeting efficiency statistics of DMY- TALENs. (H) The targeting efficiency statistics of DMY-nanos-3UTR TALENs. (I) Comparative analysis of mutation rate and germline transmission rate between DMY-TALENs and DMY-nanos3UTR-TALENs.