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In many cases, Alt-R S.p. HiFi Cas9 Nuclease 3NLS exhibits a 10–20 fold reduction in editing at problematic off-target sites, when compared with the standard Alt-R S.p. Cas9 Nuclease 3NLS. The amount of reduction in off-target editing is highly variable and can differ based on multiple factors, including off-target site, ribonucleoprotein (RNP) dose, and cell type.