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Analysis of GFP transgene integration by FACS
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When targeting the AAVS1 locus with the GFP donor18, targeted integration of GFP cDNA results in cells becoming GFP-positive, which can be easily monitored by FACS analysis. Cells were analyzed for GFP expression by flow cytometry using an Accuri C6 analyzer (BD Biosciences) 6–7 days after transfection. The relative HDR frequency was calculated by normalizing HDR frequencies by the HDR frequency induced by Cas9 alone. GFP-positive cells ranged between 0.5% and 15% of cells analyzed depending on the cell line used and the experiment.
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