High throughput gene expression profiling of yeast colonies with microgel-culture Drop-seq

Lab Chip. 2019 May 14;19(10):1838-1849. doi: 10.1039/c9lc00084d.


Yeast can be engineered into "living foundries" for non-natural chemical production by reprogramming them via a "design-build-test" cycle. While methods for "design" and "build" are relatively scalable and efficient, "test" remains a bottleneck, limiting the effectiveness of the procedure. Here we describe isogenic colony sequencing (ICO-seq), a massively-parallel strategy to assess the gene expression, and thus engineered pathway efficacy, of large numbers of genetically distinct yeast colonies. We use the approach to characterize opaque-white switching in 658 C. albicans colonies. By profiling the transcriptomes of 1642 engineered S. cerevisiae strains, we assess gene expression heterogeneity in a protein mutagenesis library. Our approach will accelerate synthetic biology by allowing facile and cost-effective transcriptional profiling of large numbers of genetically distinct yeast strains.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Gels / chemistry
  • Gene Expression Profiling
  • Genetic Engineering
  • High-Throughput Nucleotide Sequencing*
  • Particle Size
  • Saccharomyces cerevisiae / genetics*
  • Surface Properties


  • Gels