RNAi-assisted genome evolution in Saccharomyces cerevisiae for complex phenotype engineering.

摘要: A fundamental challenge in basic and applied biology is to reprogram cells with improved or novel traits on a genomic scale. However, the current ability cell genome scale limited bacterial cells. Here, we report RNA interference (RNAi)-assisted evolution (RAGE) as generally applicable method for genome-scale engineering yeast Saccharomyces cerevisiae. Through iterative cycles of creating library RNAi induced reduction-of-function mutants coupled high throughput screening selection, RAGE can continuously improve target trait(s) by accumulating multiplex beneficial genetic modifications an evolving genome. To validate constructed DNA convergent-promoter expression cassette, demonstrated cerevisiae first time identifying two known three suppressors telomerase-deficient mutation yku70Δ. We then showed application acetic acid tolerance, key trait microbial production chemicals fuels. Three rounds led identification gene knockdown targets that acted synergistically confer engineered strain substantially tolerance. should greatly accelerate design organisms desired provide new insights structure, function, evolution.