Extended Data Fig. 3: Development and validation of MPIVA.
(a-b) In vitro cleavage on (a) L3 (UA to CA mutant) and (b) SVL (CA to UA mutant) and their IC50. Compound 2 concentration used is the same as Figs. 1~2. (c) A density plot for the resistance scores of all variants in SVL-N23 library. The low, medium, and high groups represent the screens in the presence of 0.5, 2.5, and 12.5 μM Compound 2. (d) A scatter plot comparing the cleavage efficiency log(frequency in Library 2/frequency in Library 1) and the resistance score (log(frequency in Library 5/frequency in Library 2)) of SVL-CS variants. Pearson correlation is shown. (e-f) In vitro cleavage validation experiment of 4 more RNA (2 sensitive and 2 resistant) from both (e) L3-N23 and (f) SVL-N23 libraries. The CS region sequence and their IC50 is shown. (a, b) n=2 biological replicates and both measurements are shown as dots.
