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ACS Synth Biol. 2013 Dec 20;2(12):684-9. doi: 10.1021/sb400037a. Epub 2013 May 22.

Controlling mammalian gene expression by allosteric hepatitis delta virus ribozymes.

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Department of Biomedical Engineering, University of California, Davis , 451 Health Sciences Drive, Davis, California 95616, United States.


We engineered small molecule responsive allosteric ribozymes based on the genomic hepatitis delta virus (HDV) ribozyme by replacing the P4-L4 stem-loop with an RNA aptamer through a connector stem. When embedded in the 3' untranslated region of a reporter gene mRNA, these RNA devices enabled regulation of cis-gene expression by theophylline and guanine by up to 29.5-fold in mammalian cell culture. Furthermore, a NOR logic gate device was constructed by placing two engineered ribozymes in tandem, demonstrating the modularity of the RNA devices. The significant improvement in the regulatory dynamic range (ON/OFF ratio) of the RNA devices based on the HDV ribozyme should provide new opportunities for practical applications.

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