The siRNA Non-seed Region and Its Target Sequences Are Auxiliary Determinants of Off-Target Effects
Fig 5
Summary of ‘highly effective’ and ‘off-target effect-reduced’ design rules for siRNA molecules.
Based on our previous analysis and the results shown in Figs 2–4, we recommend designing molecules with: (1) A or U at position 1, (2) G or C at position 19, (3) low predicted Tm/AU richness at positions 2–7, (4) absence of GC stretches longer than 9 nt, (5) high predicted Tm in siRNA duplex/high GC content in guide strand at positions 8–15, and (6) high average GC content for all off-target sequences corresponding to positions 8–15 in the siRNA non-seed region.