Chapter 5 —— 105 —— AAVS1-targeted and precisely edited cells in the presence of ouabain was paralleled by a remarkable expansion of ATP1A1-edited cells as assessed by RFLP assays (Figure 1F, bottom panel; and Supplementary Figure S2). This data demonstrates that besides enriching for gene-targeted cell fractions in an ouabain-dependent manner, selector AAV-HRS1.A1 construct deployment is valuable for purging said fractions from off-target and/or imprecise genome editing byproducts. These results further support the robust ouabain-resistance phenotype conferred by installing the RD polymorphisms at ATP1A1. Figure 1. Testing and characterizing selector AAV genome editing at AAVS1 and ATP1A1. (A) The marker-free co-selection principle. The cell population fraction whose intracellular milieu is conducive for HR (e.g., cells undergoing the late G2/S phases of the cell cycle), are prone to simultaneous HR-mediated editing at two independent loci. As corollary, cells co-edited at a target sequence of interest and at a secondary selectable locus
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