Figure 5.

Expanding CRISPR-RNAa to enhance translation of the native E. coli gene lacZ, encoding β-galactosidase. (A) Target location of three crRNAs to enhance lacZ translation. Established transcription start sites (TSSs) are indicated, ranked in order of most common (TSS1) to least common (TSS4). (B) LacZ activity as measured using a β-galactosidase (‘Miller’) assay, performed at 30°C with 500 μM IPTG induction (as in Supplementary Figure S7, excluding step 4). (C) The same experiment, including step 4 (dilution and removal of IPTG) to enhance differences between dCasRx and dCasRx-IF3. All error bars represent standard deviation of biological triplicates. Asterisks indicate P-values of two-tailed type II Student's t-tests.