RT Journal Article
SR Electronic
T1 High throughput gene expression profiling of yeast colonies with microgel-culture Drop-seq
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 416966
DO 10.1101/416966
A1 Leqian Liu
A1 Chiraj Dalal
A1 Ben Heineike
A1 Adam Abate
YR 2018
UL http://biorxiv.org/content/early/2018/09/15/416966.abstract
AB Yeasts can be engineered into “living foundries” for non-natural chemical production by reprogramming their genome using a synthetic biology “design-build-test” cycle. While methods for “design” and “build” are scalable and efficient, “test” remains a labor-intensive bottleneck, limiting the effectiveness of the genetic reprogramming results. 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 transcriptomes of 1642 engineered S. cerevisiae strains, we use it to 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.