Advanced eMAGE for highly efficient combinatorial editing of a stable genome [article]

Zhuobin Liang, Eli Metzner, Farren J Isaacs
2020 bioRxiv   pre-print
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
more » ... multiple edits at high efficiencies in a stable genetic background, expanding utility of eMAGE in eukaryotic cells.
doi:10.1101/2020.08.30.256743 fatcat:hlofngts35bsxopu2v4g5umaoq