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. 2022 Feb 15;54(2):156–168. doi: 10.1038/s12276-022-00731-1

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a A representation of our designed CRISPR/dCas9-gRNA system whereby specific gRNAs recruit the dCas9 protein and the catalytic domain of TET1 to demethylate the targeted genomic locus. b, d, f qMSP with SW480 cells transfected with dCas9-TET1CD mock or gRNA specific to b PDX1, d EN2, and f MSX1 indicates that the designed primers can distinguish the lack of methylation modulated by the CRISPR/dCas9-gRNA system compared with controls. c, e, g qPCR with SW480 cells transfected with dCas9-TET1CD mock or gRNA of c PDX1, e EN2, and g MSX1 shows a reduction in gene expression with decreased methylation. Genomic DNA and RNA used in qMSP and qPCR were simultaneously extracted from the cell lines.