Fig. 1: CRISPR/Cas9-mediated target-specific mutations in CD34+ cells from umbilical cord blood.
From: Modeling clonal hematopoiesis in umbilical cord blood cells by CRISPR/Cas9
A Experimental timeline from cord blood to in vitro long-term culture experiments. p2/p3 depict the time points of methylcellulose replating. FACS: flow cytometry of cell surface markers. B Exemplary cell surface marker distribution of CD34+ cells at the day of transfection. C T7 endonuclease I assay of each target to determine CRISPR/Cas9 efficiency. Black arrowheads highlight the digested bands. T7E1: T7 endonuclease I, ex: exon, ctrl: Cells transfected with Cas9 protein only. D Boxplot of predicted mutation frequencies as determined by Sanger sequencing and sequence decomposition using ICE web tool. KO: insertions and deletions. SNV: hotspot mutation R882H.
