Eukaryotic multiplex genome engineering (eMAGE) offers a powerful tool to generate precise combinatorial genome modifications in Saccharomyces cerevisiae. We optimize the design of synthetic oligonucleotides and enrichment of edited populations to increase editing frequencies up to 90%, reduce workflow time by 40%, and engineer a tunable mismatch repair system to lower the rate of spontaneous mutations ~17-fold. These advances transiently evade genome maintenance to introduce multiple edits at high efficiencies in a stable genetic background, expanding utility of eMAGE in eukaryotic cells.

中文翻译：

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先进的eMAGE用于稳定基因组的高效组合编辑

真核多重基因组工程（eMAGE）提供了一个强大的工具，可以在酿酒酵母中产生精确的组合基因组修饰。我们优化了合成寡核苷酸的设计，优化了编辑后的种群，使编辑频率提高了90％，工作流程时间减少了40％，并设计了可调节的错配修复系统，将自发突变率降低了17倍。这些进展短暂地逃避了基因组维护，在稳定的遗传背景下高效导入了多个编辑，扩大了eMAGE在真核细胞中的效用。