Fig 1: Targeted deletion of the BCL11A enhancer with CRISPR-Cas9. Targeted deletion of the core sequence of the +58DHS within the erythroid-specific enhancer of the BCL11A gene using the combination of sgRNAs AC, AD, BC, and BD was confirmed by conventional PCR. PCR was performed (A) on the bulk of the KU812 cells, and (B) on the bulk of the KG-1 cells. (C) Homozygously deleted (Biallelic deletion) clones of KU812 cells, and (D) KG-1 cells. Expected band sizes for gA + gC were as ND: 920 bp, and D: 556 bp; for gA + gD were as ND: 1194 bp, and D: 658 bp; for gB + gC were as ND: 775 bp, and D: 537 bp; for gB + gD were as ND: 1049 bp, and D: 639 bp. Abbreviations: D: deleted band. ND: Non-deleted band. NC: negative control. BenchTop 100 bp DNA Ladder, Promega, Cat.: #G8291 was used as the marker. All experiments were performed twice.
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Fragment Sizes:100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1500 bp