RNAi-assisted genome evolution (RAGE) in Saccharomyces cerevisiae

Tong Si, Huimin Zhao

Research output: Contribution to journalArticle

Abstract

RNA interference (RNAi)-assisted genome evolution (RAGE) applies directed evolution principles to engineer Saccharomyces cerevisiae genomes. Here, we use acetic acid tolerance as a target trait to describe the key steps of RAGE. Briefly, iterative cycles of RNAi screening are performed to accumulate multiplex knockdown modifications, enabling directed evolution of the yeast genome and continuous improvement of a target phenotype. Detailed protocols are provided on the reconstitution of RNAi machinery, creation of genome-wide RNAi libraries, identification and integration of beneficial knockdown cassettes, and repeated RAGE cycles.

Original languageEnglish (US)
Pages (from-to)183-198
Number of pages16
JournalMethods in Molecular Biology
Volume1470
DOIs
StatePublished - Jan 1 2016

Keywords

  • Directed evolution
  • Genome engineering
  • High-throughput screening
  • Inhibitor tolerance
  • RNA interference
  • Saccharomyces cerevisiae
  • Synthetic biology

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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