A general design strategy for protein-responsive riboswitches in mammalian cells

作者：Auslaender Simon; Stuecheli Pascal; Rehm Charlotte; Auslaender David; Hartig Joerg S; Fussenegger Martin^*

来源：Nature Methods, 2014, 11(11): 1154-1160.

DOI：10.1038/NMETH.3136

RNAs are ideal for the design of gene switches that can monitor and program cellular behavior because of their high modularity and predictable structure-function relationship. We have assembled an expression platform with an embedded modular ribozyme scaffold that correlates self-cleavage activity of designer ribozymes with transgene translation in bacteria and mammalian cells. A design approach devised to screen ribozyme libraries in bacteria and validate variants with functional tertiary stem-loop structures in mammalian cells resulted in a designer ribozyme with a protein-binding nutR-boxB stem II and a selected matching stem I. In a mammalian expression context, this designer ribozyme exhibited dose-dependent translation control by the N-peptide, had rapid induction kinetics and could be combined with classic small molecule-responsive transcription control modalities to construct complex, programmable genetic circuits.

• 出版日期2014-11