RT Journal Article
SR Electronic
T1 Cooperative riboregulation in living cells through allosteric programming of toehold activation
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 009688
DO 10.1101/009688
A1 Guillermo Rodrigo
A1 Satya Prakash
A1 Shensi Shen
A1 Eszter Majer
A1 José-Antonio Daròs
A1 Alfonso Jaramillo
YR 2014
UL http://biorxiv.org/content/early/2014/09/26/009688.abstract
AB 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.