Engineering a circular riboregulator in Escherichia coli

Circular RNAs have recently been shown to be important gene expression regulators in mammalian cells. However, their role in prokaryotes remains elusive. Here, we engineered a synthetic riboregulator that self-splice to produce a circular molecule, exploiting group I permuted intron-exon (PIE) sequences. We demonstrated that the resulting circular riboregulator can activate gene expression, showing increased dynamic range compared to the linear form. We characterized the system with a fluorescent reporter and with an antibiotic resistance marker. Thanks to the increased regulatory activity by higher stability, isolation due to self-splicing, and modularity of PIE, we envisage engineered circular riboregulators in further synthetic biology applications.