High-throughput functional testing of Shiftless variants
The human protein Shiftless was recently identified as a broad-spectrum, but relatively low efficacy inhibitor of programmed ribosomal frameshifting (PRF). Here, we report results from an integrated high-throughput screening system that enables quantitative assessment of PRF inhibition by large libraries of protein variants. Using this platform, we performed deep mutagenesis of Shiftless and measured the activity of over 5,000 variants in a cell-based dual-fluorescence reporter assay with NGS-based readout.
The human protein Shiftless was recently identified as a broad-spectrum, but relatively low efficacy inhibitor of programmed ribosomal frameshifting (PRF). Here, we report results from an integrated high-throughput screening system that enables quantitative assessment of PRF inhibition by large libraries of protein variants. Using this platform, we performed deep mutagenesis of Shiftless and measured the activity of over 5,000 variants in a cell-based dual-fluorescence reporter assay with NGS-based readout.