title: Controlling mammalian gene expression by allosteric hepatitis delta virus ribozymes.
abstract: 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.
authors.affiliation: Department of Biomedical Engineering, University of California, Davis , 451 Health Sciences Drive, Davis, California 95616, United States.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Nomura
authors.forename: Linlin
authors.initials: L
authors.lastname: Zhou
authors.forename: Anh
authors.initials: A
authors.lastname: Miu
authors.forename: Yohei
authors.initials: Y
authors.lastname: Yokobayashi
mesh_terms: D001412:Bacillus subtilis; D005786:Gene Expression Regulation / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D003698:Hepatitis Delta Virus / Q000235:genetics*; D006801:Humans; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000235:genetics*; D058928:Riboswitch / Q000235:genetics; D058615:Synthetic Biology / Q000379:methods*
chemical_list: D016337:RNA, Catalytic; D058928:Riboswitch; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins