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. 2019 Nov 11;15:392–402. doi: 10.1016/j.omtm.2019.10.014

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© 2019 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Characterization of the CRISPR-Edited iPSC-RPE Cell Monolayer

(A and B) Brightfield micrograph of mature iPSC-RPE PRPF31^+/+ (A) and PRPF31^+/− (B) cells on transwells. (C and D) Fluorescent micrographs of mature CRISPR-edited iPSC-RPE PRPF31^+/+ (C) and PRPF31^+/− (D) cells grown on transwells and immunostained with anti-ZO-1 antibody (red) and DAPI (blue). (E) RPE markers normalized to UBE2R2 expressed by mature iPSC-RPE cells wild-type PRPF31^+/+ (+/+ black bars) and mutant PRPF31^+/− (+/− gray bars) compared to iPSCs (white bars, no expression found) (n = 3 replicates/cell type; data are represented as mean ± SD). (F) Transepithelial electrical resistance (TER) of mature iPSC-RPE cells on transwells (four replicates/genotype; two-way ANOVA, ***p < 0.001; data are represented as mean ± SD). (G and H) Quantification of FITC-labeled POSs (G) bound and (H) internalized by CRISPR-edited iPSC-RPE PRPF31^+/+ and PRPF31^+/− cells at 30 min, 2 h, and 5 h (two-way ANOVA, **p < 0.01, ****p < 0.0001; data are represented as mean ± SD).