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Supporting Information Fig. S1. Mutation profiles of the six3 gene found in individual embryos injected with sgRNAs targeting different regions. Top sequence shows the positions of target- 1 and 2 relative to the translation initiation codon (ATG, marked as Met) to cause frame-shifts. Note that target sites are partially overlapping. �Target 1� lists representative mutated sequences after recloning from two embryos (emb1-1 and emb1-3, see also Fig. 4f, six3 target 1) injected with sgRNA for target site 1. Eleven clones from one embryo (emb1-3) and ten clones from a second embryo (emb1-1) were analyzed. Note that no wild-type sequence (WT) was recovered. �Target 2� shows representative mutated sequences from two embryos (emb2-1 and emb2-2, see also Fig. 4f, six3 target 2) injected with sgRNA to target site 2. Nine clones from one embryo (emb2-1) and five clones from a second embryo (emb2-2) were analyzed. Note that WT sequence was recovered though at a relatively low frequency. �Proximal promoter targets� shows the -574 to +34 region with two sgRNA target sites and representative mutated sequences after recloning (14 clones) from an injected embryo (emb3, see also Fig. 4e). +1 is a tentative transcriptional start site according to the 5' end of an EST clone (accession number: DC185735). Target sequences and corresponding PAM regions are shaded in purple and red, respectively. The alignment is shown as described in Fig. 2b.

Image published in: Nakayama T et al. (2013)

Copyright © 2013. Image reproduced with permission of the Publisher, John Wiley & Sons.

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