High-resolution scanning of optimal biosensor reporter promoters in yeast

Francesca Ambri, Vasil D'ambrosio, Roberto Di Blasi, Jerome Maury, Simo Abdessamad Baallal Jacobsen, Douglas McCloskey, Michael Krogh Jensen*, Jay D. Keasling

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Small-molecule binding allosteric transcription factors (aTFs) derived from bacteria enable real-time monitoring of metabolite abundances, high-throughput screening of genetic designs, and dynamic control of metabolism. Yet, engineering of reporter promoter designs of prokaryotic aTF biosensors in eukaryotic cells is complex. Here we investigate the impact of aTF binding site positions at single-nucleotide resolution in >300 reporter promoter designs in Saccharomyces cerevisiae. From this we identify biosensor output landscapes with transient and distinct aTF binding site position effects for aTF repressors and activators, respectively. Next, we present positions for tunable reporter promoter outputs enabling metabolite-responsive designs for a total of four repressor-type and three activator-type aTF biosensors with dynamic output ranges up to 8- and 26-fold, respectively. This study highlights aTF binding site positions in reporter promoters as key for successful biosensor engineering and that repressor-type aTF biosensors allows for more flexibility in terms of choice of binding site positioning compared to activator-type aTF biosensors.
Original languageEnglish
JournalACS Synthetic Biology
Volume9
Issue number2
Pages (from-to)218-226
ISSN2161-5063
DOIs
Publication statusPublished - 2020

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