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Effect of the addition of 5′-GG to the 20 nt guide sequence on specificity 5′-GG was prefixed to the 20 nucleotide guides for the 5 most prevalent TGFBI mutations (as shown in Fig. 3a) were targeted to wild-type and respective mutant sequence in a dual luciferase assay. The addition of 5′-GG did not improve specificity, in some cases it caused a reduction in on-target activity.
