%0 Journal Article %A Guillermo Rodrigo %A Satya Prakash %A Shensi Shen %A Eszter Majer %A José-Antonio Daròs %A Alfonso Jaramillo %T Cooperative riboregulation in living cells through allosteric programming of toehold activation %D 2014 %R 10.1101/009688 %J bioRxiv %P 009688 %X Living cells rely on small non-coding RNAs (sRNAs) to regulate gene expression at the post-transcriptional level. Contrary to most protein-based activators of transcription, all known riboregulators do not exploit cooperative binding mechanisms to activate gene expression. We conceived a general strategy to design cooperative riboregulation by programming a hierarchical toehold activation cascade, which we implemented into a de novo sequence design algorithm. We engineered different riboregulatory systems relying on the conditional formation of a heterotrimeric species. We characterized the specificity of each RNA-RNA interaction in vitro and the cooperative activation of gene expression in Escherichia coli. As we only rely on a biochemical model to compute allosteric regulation, our strategy could be applied to reach more complex RNA-based nanostructures regulating gene expression for synthetic biology applications. %U https://www.biorxiv.org/content/biorxiv/early/2014/09/26/009688.full.pdf