Extended Data Fig. 3: In vitro characterization of pre-miR-155-binder and its derivatives for pre-miR-155’s A bulge.
From: Programming inactive RNA-binding small molecules into bioactive degraders
a, Representative binding curves of pre-miR-155-binder and two A bulges identified from the 2DCS selection and a fully based paired control, as determined by microscale thermophoresis (MST). The bulge found in pre-miR-155 is highlighted in green. b, Representative binding curves for pre-miR-155-binder and its derivatives for a minimized A bulge construct or fully paired mutant, as determined by MST. c, Representative binding curves for pre-miR-155-binder and its derivatives and pre-miR-155’s A bulge labelled with 2-aminopurine (2AP) or a fully base paired control (n = 2 independent experiments). d, Summary of Kds of pre-miR-155-binder analogues and other imidazolium salts identified by 2DCS. The affinity of C19 for the A bulge was undetermined due to its aggregation under assay conditions. e, Wild type pre-miR-155 but not the AU based paired mutant RNA can compete the binding of pre-miR-155-RiboTAC (5 µM) to 2AP labelled RNA (1 µM) with a Kd of 2.2 ± 0.9 μM (n = 2 independent experiments). f, pre-miR-155-RiboTAC induces in vitro RNA cleavage with wild-type (WT) pre-miR-155 construct (left), while no cleavage with mutated base pair control (right) (n = 3 independent experiments). All data are reported as the mean ± S.D. of independent replicates. All p-values were calculated using a two-tailed Student’s t-test.
