Controlling mammalian gene expression by allosteric hepatitis delta virus ribozymes

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


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis
  • Gene Expression Regulation / genetics*
  • Genetic Engineering / methods*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Hepatitis Delta Virus / genetics*
  • Humans
  • Nucleic Acid Conformation
  • RNA, Catalytic / genetics*
  • Riboswitch / genetics
  • Synthetic Biology / methods*


  • RNA, Catalytic
  • Riboswitch
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins