Modularization, minimization, and diversification of the yeast transcription factor repertoire
Authors: Daniel T. Lusk; Alessandro L.V. Coradini; Cara B. Hull; Oscar M. Aparicio; Ian M. Ehrenreich
Affiliation: University of Southern California
Keywords: Genetic interactions
Chromosome synthesis can be used to reorganize genomes in ways that help improve understanding of cellular life and evolution. Here, we are engineering a yeast strain that can be used to better explore global transcriptional control and its relationship to phenotypic diversity. We are relocating the roughly ~200 DNA binding, RNA polymerase II-associated transcription factors (TFs) in Saccharomyces cerevisiae into a single functional module on a neochromosome. The TF neochromosome is being synthesized by assembling ~260 gene-sized pieces of synthetic DNA into a single molecule. We are also constructing TF-free native chromosomes through cloning and reassembly of TF-free natural DNA segments. The outcome of this work will be a cell in which nearly all transcriptional regulation is controlled by a synthetic module on a distinct chromosome. This module will provide a platform for probing minimal sets of TFs required for viability. A minimal TF cohort will then enable explorations of the phenotypic diversity achievable through the reintroduction of accessory TFs.