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. 2018 Oct 23;1(5):337–347. doi: 10.1089/crispr.2018.0009

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© Atsushi Kunii et al. 2018; Published by Mary Ann Liebert, Inc.

This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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FIG. 4. — Robust transcriptional activation with single TREE system. (A) Assessment of the activation capacity of single TREE systems containing sgRNA #3, #4, or #5 by CDH1 reporter assay in MIA-PaCa2 cells with the comparison with the previous systems. Data are shown as the mean ± SD (n = 4). **p < 0.01; *p < 0.05. N.S., not significant. (B) Assessment of the activation capacity of single TREE systems containing sgRNA #5 by qPCR analysis of endogenous CDH1 gene in MIA-PaCa2 cells with the comparison with the previous systems. Data are shown as the mean ± SD (n = 3). *p < 0.05. N.S., not significant; N.D., not detected. (C) Assessment of the activation capacity of single TREE systems containing sgRNA #1 or #2 by RANKL reporter assay in HEK293T cells with the comparison with the previous systems. Data are shown as the mean ± SD (n = 4). **p < 0.01. (D) Assessment of the activation capacity of single TREE systems containing sgRNA #2 by qPCR analysis of endogenous RANKL gene in HEK293T cells with the comparison with the previous systems. Data are shown as the mean ± SD (n = 4). **p < 0.01; *p < 0.05. N.S., not significant. For dCas9-VPR, sgRNAs without MS2 stem loops were used instead of sgRNA2.0.