Fig. 1

Mutations in znf16-like reduce mbp mRNA expression in the CNS. (A) At 5dpf, expression of mbp was specifically reduced in the CNS but not the PNS of st78 homozygote compared with the heterozygous sibling. (B) st78 mutant had normal gross morphology at 5dpf compared with the wild-type sibling. (C) Genetic mapping linked st78 locus to XM_694039 in linkage group 7. (D) Sequence chromatogram shows the lesion (red box) in the coding sequence of znf16-like (XM_694039). The point mutation in st78 changes a cysteine in a Cys₂His₂ zinc finger of Znf16l to a phenylalanine. (E) Schematic diagram depicting Znf16-like protein with all its zinc fingers, the location of the transversion in the st78 allele, and the 4bp deletion resulting in a frameshift in the st97 allele. (F) Nucleotide sequence of znf16l and the TALE nuclease targeted sequence (black) and diagnostic restriction enzyme site (red). Bottom line shows the st97 allele and the corresponding amino acid sequence resulting from the frameshift. (G) At 5dpf, expression of mbp was reduced in the CNS of st97 homozygote in comparison to the heterozygous sibling. st97 failed to complement st78, and the transheterozygous mutant lacked mbp in the CNS, similar to the homozygous mutants. The genotypes of all larvae shown in A,B,G,H were determined by PCR tests for their respective lesions.