A universal system for boosting gene expression in eukaryotic cell-lines

Inbal Vaknin, Or Willinger, Jonathan Mandl, Hadar Heuberger, Dan Ben-Ami, Yi Zeng, Sarah Goldberg, Yaron Orenstein, Roee Amit

Research output: Contribution to journalArticlepeer-review

Abstract

We demonstrate a transcriptional regulatory design algorithm that can boost expression in yeast and mammalian cell lines. The system consists of a simplified transcriptional architecture composed of a minimal core promoter and a synthetic upstream regulatory region (sURS) composed of up to three motifs selected from a list of 41 motifs conserved in the eukaryotic lineage. The sURS system was first characterized using an oligo-library containing 189,990 variants. We validate the resultant expression model using a set of 43 unseen sURS designs. The validation sURS experiments indicate that a generic set of grammar rules for boosting and attenuation may exist in yeast cells. Finally, we demonstrate that this generic set of grammar rules functions similarly in mammalian CHO-K1 and HeLa cells. Consequently, our work provides a design algorithm for boosting the expression of promoters used for expressing industrially relevant proteins in yeast and mammalian cell lines.

Original languageEnglish
Article number2394
JournalNature Communications
Volume15
Issue number1
DOIs
StatePublished - 1 Dec 2024

ASJC Scopus subject areas

  • General Chemistry
  • General Biochemistry, Genetics and Molecular Biology
  • General Physics and Astronomy

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