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Insertion of the GBT cassette into the second intron of zebrafish kcng4b causes a mutation. (A-D) Homozygous mutants show a clump of cells in the third ventricle (30 hpf, B,D,D′, white arrow) and hydrocephalus (48 hpf, F,G, black arrow and arrowheads). (H) TAIL-PCR mapped the insertion at position 57,917,547 on Chr. 7 in the (+) orientation (Zv9). (I) Insertion into the kcng4b locus truncates the full-length transcript. kcng4b−/− expresses the truncated transcript-kcng4b* consisting of one coding exon. (J) RT-PCR analysis demonstrates a reduction of kcng4b transcripts in kcng4b+/− and their absence in kcng4b−/− (30 hpf). Actin was detected as a loading control. (A,B) Frontal views, (C-G) lateral views, anterior to the left. E, exon; wt, wild type; A and B, primers as detailed in the Materials and Methods.
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