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Fig. 1 CRISPR-Cas9 mediated Aqp0a or Aqp0b functional knock-out. (A) gRNA designed against exon 1 of aqp0a or aqp0b co-injected with Cas9 resulted in out-of-frame deletions (11 bp for aqp0a and 25 bp for aqp0b) that are predicted to create premature stop codons. Mutants identified by PCR were confirmed by DNA sequencing. (B) Maternal-zygotic mutant adult lens fiber membrane protein was run on a Western blot. (B, top) Failure to detect at the monomeric molecular weight of ∼28 kDa (arrow), as well as labeling of the polymeric bands was observed in aqp0a−/− by anti-Aqp0a antibody, and aqp0b−/− by anti-Aqp0b antibody, while the other ortholog was still detected. Stripping and reprobing the blot with anti-α-Tubulin antibody (B, bottom) served as a protein loading control.