Synthetic Translational Regulation by Protein-Binding RNA Origami Scaffolds

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Abstract

Rational design approaches for the regulation of gene expression are expanding the synthetic biology toolbox. However, only a few tools for regulating gene expression at the translational level have been developed. Here, we devise an approach for translational regulation using the MS2 and PP7 aptamer and coat-protein pairs in Escherichia coli. The aptamers are used as operators in transcription units that encode proteins fused to their cognate coat proteins, which leads to self-repression. RNA origami scaffolds that contain up to four aptamers serve as an alternate binder to activate translation. With this system, we demonstrate that the increase in expression of a reporter protein is dependent on both the concentration and number of aptamers on RNA origami scaffolds. We also demonstrate regulation of multiple proteins using a single MS2 coat protein fusion and apply this method to regulate the relative expression of enzymes of the branched pathway for deoxyviolacein biosynthesis.

Original languageEnglish
JournalA C S Synthetic Biology
Volume11
Issue5
Pages (from-to)1710-1718
Number of pages9
ISSN2161-5063
DOIs
Publication statusPublished - 20 May 2022

Keywords

  • RNA origami
  • RNA synthetic biology
  • RNA-protein complexes
  • post-transcriptional regulation

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