Figure 3.

CRISPR-engineered IPO7 mutants in 721 cells. 721 cells were transfected with guide RNAs that targeted exon1 (A), or exon4 (B) along with Cas9, as described in Figure 2. The wild-type sequence is shown; the target sequence is shown in red; and PAM is underlined and shown in green. The sequence of each clone was determined through sequencing and the mutated genomic sequence and its encoded amino acids are shown in blue. (C) The IPO7 expression of certain IPO7 mutants was decreased as detected via Western blotting. The expression of α-Tubulin was also determined as an internal control. The mean intensity of IPO7 in these clones was 0.22 relative to the control with a S.E. +/−0.07.