title: RNAi-Assisted Genome Evolution (RAGE) in Saccharomyces cerevisiae.
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.
authors: Carl R Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
authors: Tong
authors: T
authors: Si
authors: Carl R Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA. zhao5@illinois.edu.|Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA. zhao5@illinois.edu.|Departments of Chemistry, Biochemistry, and Bioengineering, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, IL, 61801, USA. zhao5@illinois.edu.
authors: Huimin
authors: H
authors: Zhao
mesh_terms: D019342:Acetic Acid / Q000378:metabolism / Q000494:pharmacology; D019020:Directed Molecular Evolution / Q000379:methods*; D055785:Gene Knockdown Techniques / Q000379:methods; D015723:Gene Library; D005818:Genetic Engineering / Q000379:methods*; D016681:Genome, Fungal; D057166:High-Throughput Screening Assays / Q000379:methods; D034622:RNA Interference; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism
chemical_list: D019342:Acetic Acid
keywords: Directed evolution; Genome engineering; High-throughput screening; Inhibitor tolerance; RNA interference; Saccharomyces cerevisiae; Synthetic biology