Fig. 2: Pre-miR-155-RIBOTAC selectively degrades pre-miR-155 in an RNase-L-dependent manner in breast cancer cells.
From: Programming inactive RNA-binding small molecules into bioactive degraders
a, Schematic of converting an inert binder engaging pre-miR-155 into an active RIBOTAC degrader. b, Structures of the compounds used to target pre-miR-155. c, The effects of pre-miR-155-RIBOTAC on mature (mat) (n = 4 biological replicates), pre- (n = 3 biological replicates) and pri- (n = 3 biological replicates) miR-155 levels, competed by increasing concentrations of pre-miR-155-binder in MDA-MB-231 cells. d, The effect of siRNA knockdown of RNase L on pre-miR-155-RIBOTAC-mediated cleavage of pre-miR-155 in MDA-MB-231 cells, as determined using RT–qPCR. n = 3 biological replicates. e, Immunoprecipitation of pre-miR-155 using an anti-RNase L antibody in the presence of pre-miR-155-RIBOTAC in MDA-MB-231 cells (n = 3 biological replicates). f, The effect of pre-miR-155-amide-binder (left; 100 nM; n = 4 biological replicates) and pre-miR-155-RIBOTAC (right; 100 nM; n = 3 biological replicates) on the levels of the 373 miRNAs expressed in MDA-MB-231 cells. FC, fold change. g, Western blot analysis of SOCS1, a direct target of miR-155, after treatment of MDA-MB-231 cells with pre-miR-155-RIBOTAC (n = 3 biological replicates). h, The effect of pre-miR-155-RIBOTAC on the activity of a SOCS1 3′ UTR-luciferase reporter transfected into HEK293T cells, establishing both dose (left) and time dependence (right; n = 4 biological replicates). Data are mean ± s.d. (c–e, g and h). Statistical significance was determined using two-tailed Student’s t-tests (c–e, g and h) or a Wald’s test (f).
