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. 2022 Mar 2;10:830432. doi: 10.3389/fcell.2022.830432

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Copyright © 2022 An, Kuo and Tang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Generation of CRISPR/Cas9-mediated homozygous point mutation in the CPAP/CENPJ gene. (A) The scheme of homologous recombination between endogenous CPAP wild-type (WT) and exogenous repair template (single strain DNA, ssDNA) harbored A>T point mutation. Black inverted triangle: Cas9-cutting site. (B) Sanger sequencing results of two independent mutant clones displaying homozygous A>T point mutation in exon 16 of CPAP, which resulted in Glutamic acid (E) being replaced by Valine (V) at amino acid 1235. (C) CPAP protein expression level in WT and two independent mutant hiPSC clones. Black star symbol indicated non-specific bands.