Figure 1.

High-throughput quantification of tRNA function of SUP4oc variants. (A) Schematic of the RNA-ID reporter used to quantify tRNA function. (B) SUP4_oc efficiently suppresses GFP_oc. Scatter plot of flow cytometry of cells with integrated RNA-ID reporter expressing GFP (green), GFP_oc (red), and GFP_oc and SUP4_oc (blue). (C) FACS of SUP4_oc variant library. Cells were grown in YP galactose medium and sorted. (D) SUP4oc tolerates numerous mutations. Cloverleaf heat map showing GFP^SEQ of single-mutant variants. Quadrant color around residues indicates variant activity. Active variants are white (GFP^SEQ of 0.026) to blue (GFP^SEQ of 1) gradient, and inactive variants are red. Modified bases are indicated in the figure.