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. Author manuscript; available in PMC: 2018 Apr 13.
Published in final edited form as: Nature. 2016 Nov 7;539(7629):384–389. doi: 10.1038/nature20134

Fig 2. Enrichment of HBB-targeted HSPCs using FACS and magnetic bead-based technologies.

Fig 2

a) Left panel: representative FACS plots highlight the GFPhigh population (red gate) generated by the addition of Cas9 RNP. Right panel: HBB-targeted HSPCs from GFPhigh (red), GFPlow (green), and GFPneg (blue) fractions were sorted and monitored for GFP expression. Error bars represent S.E.M. (N=11, all from unique mPB or CB donors) b) PCR was performed on methylcellulose colonies from GFPhigh HSPCs to detect targeted integration at the 3’ end.c) Left panel: representative FACS plots highlight the tNGFRhigh population (red gate) generated by the addition of Cas9 RNP. Right panel: tNGFRhigh (red) HSPCs were enriched using anti-CD271 (LNGFR) magnetic microbeads and cultured for 18 days while monitoring tNGFR expression. Error bars represent S.E.M. (N=5, all from unique CB donors). d) PCR was performed on tNGFRhigh-derived methylcellulose colonies to detect targeted integrations at the 5’ end.