TALENs-Assisted Multiplex Editing for Accelerated Genome Evolution to Improve Yeast Phenotypes

Guoqiang Zhang, Yuping Lin, Xianni Qi, Lin Li, Qinhong Wang, Yanhe Ma

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Genome editing is an important tool for building novel genotypes with a desired phenotype. However, the fundamental challenge is to rapidly generate desired alterations on a genome-wide scale. Here, we report TALENs (transcription activator-like effector nucleases)-assisted multiplex editing (TAME), based on the interaction of designed TALENs with the DNA sequences between the critical TATA and GC boxes, for generating multiple targeted genomic modifications. Through iterative cycles of TAME to induce abundant semirational indels coupled with efficient screening using a reporter, the targeted fluorescent trait can be continuously and rapidly improved by accumulating multiplex beneficial genetic modifications in the evolving yeast genome. To further evaluate its efficiency, we also demonstrate the application of TAME for significantly improving ethanol tolerance of yeast in a short amount of time. Therefore, TAME is a broadly generalizable platform for accelerated genome evolution to rapidly improve yeast phenotypes.

Original languageEnglish (US)
Pages (from-to)1101-1111
Number of pages11
JournalACS Synthetic Biology
Volume4
Issue number10
DOIs
StatePublished - Oct 16 2015

Keywords

  • cellular phenotypes
  • genome evolution
  • multiplex editing
  • transcription activator-like effector nucleases

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