Codon-restrained method for both eliminating and creating intragenic bacterial promoters

Dominic Y. Logel, Ellina Trofimova, Paul R. Jaschke*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Future applications of synthetic biology will require refactored genetic sequences devoid of internal regulatory elements within coding sequences. These regulatory elements include cryptic and intragenic promoters, which may constitute up to a third of the predicted Escherichia coli promoters. The promoter activity is dependent on the structural interaction of core bases with a σ factor. Rational engineering can be used to alter key promoter element nucleotides interacting with σ factors and eliminate downstream transcriptional activity. In this paper, we present codon-restrained promoter silencing (CORPSE), a system for removing intragenic promoters. CORPSE exploits the DNA-σ factor structural relationship to disrupt σ70 promoters embedded within gene coding sequences with a minimum of synonymous codon changes. Additionally, we present an inverted CORPSE system, iCORPSE, which can create highly active promoters within a gene sequence while not perturbing the function of the modified gene.

Original languageEnglish
Pages (from-to)689-699
Number of pages11
JournalACS Synthetic Biology
Volume11
Issue number2
Early online date19 Jan 2022
DOIs
Publication statusPublished - 18 Feb 2022

Keywords

  • refactoring
  • transcription initiation
  • internal regulation
  • synthetic genomics
  • σ factor
  • sigma factor

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