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. 2019 Aug 20;47(19):e120. doi: 10.1093/nar/gkz713

Figure 5.

Figure 5.

Highly efficient editing in human pluripotent stem cells (hPSCs) using TREE (A) Quantification of base editing efficiency (percentage GFP-positive cells) when hPSCs were co-transfected with pEF-BFP, sg(BG) and various base editing vectors. n = 3; * = P < 0.05, ** = P < 0.01. (B) Schematic for enrichment of edited hPSC using TREE. HPSCs were co-transfected with pEF-BFP, pEF-BE4-Gam/pEF-AncBE4 and pDT-sgRNA vectors. 48 h post-transfection, flow cytometry was used to sort cell populations into GFP-positive and -negative fractions. (C) Representative flow cytometry plots of (i) untransfected hPSCs cells and (ii) hPSCs transfected with pEF-BFP only as well as hPSCs cells in which TREE was applied targeting Site-1 utilizing (iii) pEF-BE4-Gam or (iv) pEF-AncBE4. (D) Quantification of base editing efficiency at Site-1 in GFP-positive, GFP-negative and unsorted cell populations isolated using TREE- or RoT-based enrichment strategies in which pEF-BE4-Gam or pEF-AncBE4 was employed. n = 3; * = P < 0.05, ** = P < 0.01.