Precise control of SCRaMbLE in synthetic haploid and diploid yeast

Nat Commun. 2018 May 22;9(1):1933. doi: 10.1038/s41467-018-03084-4.

Abstract

Compatibility between host cells and heterologous pathways is a challenge for constructing organisms with high productivity or gain of function. Designer yeast cells incorporating the Synthetic Chromosome Rearrangement and Modification by LoxP-mediated Evolution (SCRaMbLE) system provide a platform for generating genotype diversity. Here we construct a genetic AND gate to enable precise control of the SCRaMbLE method to generate synthetic haploid and diploid yeast with desired phenotypes. The yield of carotenoids is increased to 1.5-fold by SCRaMbLEing haploid strains and we determine that the deletion of YEL013W is responsible for the increase. Based on the SCRaMbLEing in diploid strains, we develop a strategy called Multiplex SCRaMbLE Iterative Cycling (MuSIC) to increase the production of carotenoids up to 38.8-fold through 5 iterative cycles of SCRaMbLE. This strategy is potentially a powerful tool for increasing the production of bio-based chemicals and for mining deep knowledge.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Carotenoids / biosynthesis*
  • Chromosomes, Fungal / chemistry
  • Clone Cells
  • Gene Deletion
  • Gene Expression Regulation, Fungal*
  • Genes, Synthetic
  • Genome, Fungal*
  • Integrases / genetics
  • Integrases / metabolism
  • Metabolic Engineering / methods*
  • Metabolic Networks and Pathways / genetics
  • Phenotype
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Ploidies*
  • Recombination, Genetic
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Vesicular Transport Proteins / deficiency
  • Vesicular Transport Proteins / genetics

Substances

  • Saccharomyces cerevisiae Proteins
  • VAC8 protein, S cerevisiae
  • Vesicular Transport Proteins
  • Carotenoids
  • Cre recombinase
  • Integrases