The scn1labΔ44 stable knockout line was generated using zinc-finger nucleases (ZFNs). A CompoZr ZFN was designed by Sigma-Aldrich to target exon 4 of scn1lab. CompoZr ZFN contained a DNA-binding domain and an obligate-heterodimer Fok1 nuclease domain, engineered for improved specificity (Miller et al., 2007 (link)). Activity of ZFN pairs as determined by the yeast MEL-1 reporter assay (Doyon et al., 2008 (link)) was 113.6%. ZFNs were prepared and used as previously described (Hoffman et al., 2016 (link)). We confirmed by Sanger sequencing the presence of a 44-nucleotide deletion in scn1lab exon 4 (chr6:10,299,906–10,299,949) and two SNPs: T>A at chr6:10,299,903 and T>C at chr6:10,299,904 (danRer11). The deletion includes the intron/exon four boundary.
ZFNs binding sequences and sequences of the PCR primers used for sequencing and genotyping are provided inSupplementary file 1 .
ZFNs binding sequences and sequences of the PCR primers used for sequencing and genotyping are provided in
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