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Fig. 6 IF at top three predicted off-target locations. IF was determined for Alt-R HDR components with 200 pg (col1a2 c.918A>T), 400 pg (col1a2 c.1416A>C, col1a2 c.2093G>A) or 800 pg (atp6v1e1b c.634_636CGA>TGG, atp6v1e1b c.383T>C, brca2 c.6803G>A) Cas9 or PE components (RNP mixture containing 480 ng/µL pegRNA and 1,380 ng/µL PE2-His protein) injected into the yolk of one-cell-stage zebrafish embryos. The bars and error bars represent the mean + s.e.m. of three biologically independent replicates (n = 3), where each replicate represents a pool of 20–30 injected embryos. Unpaired two-tailed t-test: **P < 0.01, ***P < 0.001.