A General Design Strategy for Protein-responsive Riboswitches in Mammalian Cells

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2014 Oct 6

PMID 25282610

Citations 47

Authors

Simon Auslander

Pascal Stucheli

Charlotte Rehm

David Auslander

Jorg S Hartig

Martin Fussenegger

Affiliations

Soon will be listed here.

Abstract

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.

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