Excision of H3K4me3/DNaseI signatures by dual guideRNA CRISPR vectors.

<p><b>A)</b> Primary human monocyte DNase- and ChIP-Seq (H3K4me3 and K3K27me3) coverages at the transcriptional start sites of the MALAT1, miR-146a and miR-155 ncRNA genes. Regions selected for targeted excision are marked (dashed lines). <b>B)</b> Genomic PCR with primers flanking the respective H3K4me3/DNaseI element to be excised. Besides the wild-type band (WT) a band of reduced size (ΔTSS) is detected after transfection of HEK293 cells with the respective ncRNA knockout CRISPR construct (KO) but not after transfection of a control CRISPR construct (Ctrl.). Position of the DNA ladder bands (2.5 kb, 2 kb, 1.5 kb, 1 kb, 750 bp, 500 bp, 250 bp) is indicated to the left of each gel image. <b>C)</b> Sanger-sequencing of ΔTSS bands validates the deletion in the MALAT1, miR155 and miR146a promoter, respectively. Positions of guideRNAs (gRNA) and protospacer-adjacent motifs (PAM) are indicated.