Zinc-finger nuclease-mediated targeting of zebrafish rest. (A) Sequence chromatograms from wild-type siblings (left) and rest mutants (right). The 7 bp deletion (Δ7) in SBU29 and 4 bp insertion (+4) in SBU34 are illustrated beneath. (B) Domain structure of wild-type Rest and truncated proteins produced by SBU29 and SBU34 mutations. (C) Animal pole views of 6-hpf wild-type and MZrestsbu29/sbu29 embryos. Rest immunoreactivity, TOPRO-3 staining of nuclei, and merged channels are shown. Abundant cytoplasmic Rest protein is detected in wild-type embryos, but not in MZrestsbu29/sbu29 mutants. Confocal images are single 1-μm stacks taken at 40× magnification.
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