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. 2023 Feb 8;24(4):3417. doi: 10.3390/ijms24043417

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© 2023 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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Generation of a stable CRISPRi human RPE cell line expressing dCas9-KRAB. Characterization of ARPE19-KRAB cell line showing expression of (A) KRAB by RT-PCR, and (B) SpCas9 by immunocytochemistry. (C) ARPE19-KRAB was able to form polarised monolayer upon nicotinamide treatment. Left panel shows representative morphology of ARPE19-KRAB treated with control condition (10% FCS) or differentiation media with nicotinamide. Middle and right panel show immunocytochemistry of tight junction protein ZO-1. (D) Heatmap of short tandem repeat analysis of 10 polymorphic markers. Alleles 1 and 2 are designated as −1 and −2 respectively.