Xenbase Image ID: 129444

Fig. 4. Disruption of restriction enzyme recognition sites between the left and right target sites in embryos injected with TALEN mRNAs.(A,B) Schematic drawing of the genomic PCR product containing Tyr I (A) or Tyr II (B) target sites. An HaeIII site (Hae) and a PflMI site (Pfl) are located in the spacer sequences of Tyr I and Tyr II, respectively. The primer sets and target sites are indicated as blue arrowheads and red bars, respectively. (C–F) Quantification of resistance to restriction enzyme digestion. Embryos were injected with 400 pg, 80 pg or 0 pg (control) of TALEN-Tyr I mRNAs (C,D) or TALEN-Tyr II mRNAs (E,F). Genomic DNA was separately prepared from randomly selected embryos at NF-stage 35/36 (Fig. 2; Fig. 3) and subjected to PCR using a specific primer set to amplify DNA fragments containing the target sites. The PCR products were digested with HaeIII (C,D) or PflMI (E,F) enzymes and separated on agarose gels. The injected TALEN scaffolds are shown at the bottom. The number of analyzed embryos is shown at the top of each column. Due to embryo death, genomic DNA was not extracted from embryos injected with 400 pg of δNδC-Tyr II mRNAs (E). The statistical significance compared to the control (a) or embryos injected with δNδC-ELD/KKR mRNAs (b) was assessed using a Tukey test. P<0.05. The data are expressed as the means ± s.e.m. 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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