Figure S1.

Energy Penalty of SNVs and Modeling of SNIPR Performance, Related to Figure 1

(A) Analysis of energy penalties caused by different types of SNVs. In the boxplot, the top of the rectangle indicates the third quartile, the horizontal line within the rectangle indicates the median, and the bottom of the rectangle indicates the first quartile. The upper vertical line extends to the highest data point within 1.5-fold of the interquartile range (IQR) of the top quartile, and the lower vertical line extends to the lowest data point within 1.5-fold of the IQR of the lowest quartile.

(B-C) The complex in the ON state with the WT HFE mRNA target (B) and the conformation of the complex in the OFF state with a mutant HFE C282Y target (C). Both panels show the equilibrium probability of the bases being in the indicated hybridization state. Red boxes indicate the location of the single-nucleotide sequence difference.

(D-G) Simulated kinetic curve of protein expression level with the correct target and SNV target under different reaction energies. From the model, the optimal free energy range of ΔG = −2 to −1 kcal/mol was obtained, which provided the best combination of protein expression and discrimination capacity.

(H) The relationship between the differentiation factor and the reaction energy upon binding to the correct target. The red dashed line indicates a differentiation factor of 20.