Fig. 3 - Supplemental 1

CRISPR/Cas9-induced mutation in zebrafish aqp1a.1 gene. (A) Zebrafish aqp1a.1 gene structure, gRNA binding site (in blue), aqp1a.1rk28 allele, Aqp1a.1 wildtype (261 aa) and Aqp1a.1rk28 (truncated at 76 aa) protein structure. The rk28 mutation causes a 14-nt deletion and an 8-nt insertion (in red), which leads to a premature termination codon (underlined), and as a result, to the loss of four out of six transmembrane domains/helices (H4/TMD3-H8/TMD6) and two membrane-inserted non-membrane-spanning helices (H3 and H7). (B) Sequence read showing 14-nt deletion (shadowed area in wildtype allele) and 8-nt insertion (shadowed area in rk28 allele) in aqp1a.1rk28 allele. Premature termination codon is underlined. Amino acid sequence is shown above the nucleotide sequences. (C) qPCR analysis of aqp1a.1 mRNA expression in wildtype, aqp1a.1rk28/rk28 and aqp8a.1rk29/rk29 embryos. (D) Representative images of wildtype, heterozygote, and homozygote embryos after whole-mount in situ hybridization with aqp1a.1 RNA probe at 30 hpf. Scale bar, 250 µm.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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