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Creation and validation of the fmr1 mutants using TALEN. (A) Schematic representation of the zebrafish fmr1 with transcript ENSDART00000123434.3 (15 exons, size: 4413 bp). The data were obtained from Ensembl https://asia.ensembl.org/Danio_rerio/Transcript/Summary?db=core;g=ENSDARG00000037433;r=14:20156477-20191281;t=ENSDART00000123434. The lower panel shows the TALEN constructs. Exon 1 of the fmr1 gene was targeted with the 14-bp spacer (red, underlined) flanked by TALEN arms (blue and green). (B) WT and mutant sequences. The 11 deleted amino-acids (AA) are left blank in the mutant sequence to indicate the comparison with WT. The three following AA in the WT were substituted by AAA (in red) in the mutant sequence. (C) Genotyping data showing the WT and mutants alleles. Highlighted in yellow, the mutation site. Note after TCA, the deletion of 11 amino-acids (Δ11) and the substitution of 3 amino-acids by AAA. Highlighted in light blue, the unchanged regions. (D) Western blot analysis of the adult brains.
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