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. 2022 Jun 27;14(13):3148. doi: 10.3390/cancers14133148

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Effects of optimizing the TOP2α intron 19 5′ SS in K/VP.5 cells by CRISPR/Cas9/HDR editing. (A) RNA-seq genome coverage tracks for TOP2α showing the intron 19 IPA event in K/VP.5 cells and the restoration of intron removal in K/VP.5/edit-3 cells. Reduced coverage in exon 20 in K/VP.5 cells is consistent with fewer full-length TOP2α/170 reads while greater exon 20 coverage in K/VP.5/edit-3 cells is consistent with intron 19 removal and more full-length mature TOP2α/170 mRNA. (B) Immunoassay (N-terminal specific TOP2α antibody; GAPDH antibody) using K562, K/VP.5, and KVP.5/edit-3 cellular lysates. (C) Forty-eight-hour growth inhibitory effects of etoposide in K562, K/VP.5, and K/VP.5/edit-3 cells. Republished with permission of the American Society for Pharmacology and Experimental Therapeutics, from Hernandez et al. Mol Pharmacol. 99:226–241, 2021 [23].