A precisely adjustable, variation-suppressed eukaryotic transcriptional controller to enable genetic discovery.

ABSTRACT: Conditional expression of genes and observation of phenotype remain central to biological discovery. Current methods enable either on/off or imprecisely controlled graded gene expression. We developed a 'well-tempered' controller, WTC₈₄₆, for precisely adjustable, graded, growth condition independent expression of genes in Saccharomyces cerevisiae. Controlled genes are expressed from a strong semisynthetic promoter repressed by the prokaryotic TetR, which also represses its own synthesis; with basal expression abolished by a second, 'zeroing' repressor. The autorepression loop lowers cell-to-cell variation while enabling precise adjustment of protein expression by a chemical inducer. WTC₈₄₆ allelic strains in which the controller replaced the native promoters r

SUBMITTER: Azizoglu A 

PROVIDER: S-EPMC8421071 | biostudies-literature | 2021 Aug

REPOSITORIES: biostudies-literature