Fig. 3: Characterization of f-MAGE and f-CRISPR performance.
From: Targeted editing and evolution of engineered ribosomes in vivo by filtered editing
a f-CRISPR was performed on genomic oRiboT-Tt2 (Supplementary Fig. 3) to introduce dsDNA to replace a 832-bp region of oRiboT and introduce a 7 N mutagenic library; editing-efficiency was determined by deep sequencing. The left axis quantifies the percent of oRiboT or WT ribosomes that were edited (% edited). The red dot corresponds to the rightmost axis, which quantifies the library complexity. b f-MAGE was performed with ssODNs having 0–70 nucleotides of homology to the intron, in order to determine the optimum ssODN design parameters for targeted mutagenesis and reduction of off-target mutations. The percent of oRiboT or WT ribosomes that were edited (% edited) was determined with deep sequencing.
