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. 2019 Aug 14;3(10):1356–1372. doi: 10.1002/hep4.1413

Figure 6.

Figure 6

OLMALINC enhancer/promoter deletion using CRISPR‐Cas9 gene editing decreases SCD gene expression. (A) Schematic of primer designs for genomic PCR amplification of wild type versus CRISPR‐Cas9‐mediated OLMALINC promoter/enhancer deletion. Per ENCODE HepG2 chromatin state data, red highlights OLMALINC promoter while yellow highlights the enhancer. (B) Gel electrophoresis of PCR products from amplification of the wild type and CRISPR‐Cas9 OLMALINC enhancer/promoter deletions from the genomic DNA from HepG2 cells. (C) Evaluation of transfection efficiency of HepG2 with fluorescently labeled tracRNA with ATTO‐550 after 24 hours; left panel demonstrating bright field cells and right panel the corresponding labeled cells. (D) OLMALINC and SCD gene expression by RT‐qPCR after 48‐hour transfection with the Cas9 enzyme and OLMALINC gRNAs flanking the enhancer/promoter region. Values are mean ± SD (n = 3). **P < 0.01, ***P < 0.001 (unpaired Student t test was used for two groups). Abbreviations: bp, base pair; tracRNA, trans‐activating RNA.