Fig. 1 | Nature Communications

Fig. 1

From: Gene correction for SCID-X1 in long-term hematopoietic stem cells

Fig. 1

In vitro, medium scale genome targeting at IL2RG locus. a Diagram of genomic integration and correction outcomes. b Top: schematic of IL2RG corrective donors containing (+tNGFR) or not (–tNGFR) selectable marker. Bottom: IL2RG cDNA targeting frequencies of frozen mobilized peripheral blood CD34+HSPCs (white circles) or freshly purified cord blood male-derived CD34+HSCPs (red circles) derived from medium scale (1.0 × 10 6) genome targeting and measured at day 4. Absolute targeting frequencies measured by ddPCR. Median: 23.2% (+tNGFR, n = 11 biological replicates), median 45% (–tNGFR, n = 13 biological replicates). c Single cell-based methylcellulose assay from mock targeted (nucleofected only) or IL2RG cDNA targeted (–tNGFR donor) CD34+HSPCs. Absolute number of clones are shown (n = 3 biological replicates). d Fraction of the total for each type of colony scored. e Gene correction outcome of SCID-X1 patient 2 derived CD34+HSPCs. Shown is the multi-lineage differentiation using OP9-idll1 in vitro system (n = 23 wells). No growth was derived from uncorrected CD34+cells. LT-HSPCs long-term hematopoietic stem cells, ST-HSC short term hematopoietic stem cells, MPP multi-potent progenitor, CMP common myeloid progenitor, LMPP lymphoid multi-potent progenitor, CLP common lymphoid progenitor, HSPCs hematopoietic stem and progenitor cells, ddPCR droplet digital digital droplet PCR. Mean ± s.e.m.; ns not specific (Welch’s t-test). Source data are available in the Source Data file