The results from the designed experiment to unravel the principles for translation inhibiting RNA (tiRNA) design.

(A) The practical execution of the design of experiments (DOE). All tiRNAs, representing a data point in the DOE, are coexpressed with the target untranslated region (UTR) and the riboregulator efficiency is determined. (B) All tiRNAs with corresponding feature data point in the DOE, where the features are calculated using UTR1. (C) The tiRNA performance, expressed in relative protein expression, where lower expression represents more effective tiRNAs. The performance was evaluated using both UTR1 and UTR2. The 100% relative protein expression represents the protein expression in absence of the tiRNA. Error bars represent standard deviation (n = 3).