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Fig. 5. Target site sequences in TALEN-mRNA-injected embryos.Target DNA sequences were determined using pooled genomic DNAs purified from NF-stage 35/36 embryos that had been injected with 400 pg of TALEN-Tyr I (A) or –Tyr II (B) mRNAs. (A) Mutation of target site sequences by TALEN-Tyr I. Genomic DNA was extracted from each of 16, 9, 13 and 13 embryos injected with TAL-Tyr I, ΔNΔC-Tyr I, ΔNΔC-ELD/KKR-Tyr I and ΔNΔC-ELD-S/KKR-S-Tyr I mRNAs, respectively, and pooled. (B) Mutation of target site sequences by TALEN-Tyr II. Genomic DNA was extracted from each of 7, 12 and 9 embryos injected with TAL-Tyr II, ΔNΔC-ELD/KKR-Tyr II and ΔNΔC-ELD-S/KKR-S-Tyr II mRNAs, respectively, and pooled. Due to embryo death, genomic DNA was not purified from embryos injected with ΔNΔC-Tyr II mRNAs. The wild-type sequence is shown as WT. The black bars indicate the Tyr I (A) and Tyr II (B) target sites. Gaps resulting from deletion are denoted as dashes. Inserted nucleotides are indicated as red characters. The HaeIII (A) and PflMI (B) recognition sequences are indicated as blue characters. The mutation types and frequencies are indicated on the right. (C,D) Percentage of mutant target site sequences derived from embryos injected with TALEN-Tyr I (C) or TALEN-Tyr II (D) mRNAs. Image published in: Nakajima K and Yaoita Y (2013) © 2013. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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