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. 2019 Aug 5;15(8):e1008319. doi: 10.1371/journal.pgen.1008319

Fig 2. RMR frequencies in parental U2OS cells.

Fig 2

(A) Repeat-mediated repair (RMR) reporter diagram. The GFP coding sequence is disrupted by a non-homologous insert, and for each reporter the 3' GFP fragment shares homology with the 5' GFP fragment, but with varying lengths of homology (200–6 nt). Each reporter was integrated into cells using the FRT/Flp system. (B) GFP+ repair events induced by an sgRNA/Cas9-mediated DSB targeted to the 5' edge of the non-homologous insert (5' edge). Shown is the GFP+ frequency induced by the 5' edge DSB for each reporter cell line with varying lengths of homology in the 3' GFP fragment (200–6 nt), each normalized to transfection efficiency. Two independent clones were tested for each reporter with four independent replicates, for a total of n = 8, except for the 23 nt reporter where six independent clones were tested for a total of n = 24. Error bars represent SD. (C) GFP+ repair events induced by expressing sgRNAs with Cas9 that cause DSBs at the 5' edge of the non-homologous insert (5' edge), the 3' edge of the non-homologous insert (3' edge), or both the 5' and 3' edge of the non-homologous insert (5' & 3' edge). Shown is the percentage of GFP+ cells for each reporter cell line with ≤ 50 nt of homology in the 3' GFP fragment (50–6 nt), which was normalized to transfection efficiency. Error bars represent SD. The number of clones tested and total n as in (B).