title: Programming a highly structured ribozyme into complex allostery using RNA oligonucleotides.
abstract: RNA possesses great potential for expanding the toolbox currently available to synthetic biologists. Here, the modulation of the Hepatitis Delta Virus ribozyme's activity with a series of rationally designed aptamers and effector RNA oligonucleotides is described. The ribozyme was initially fused with an 18-nucleotide hairpin structure that abolished its self-cleaving activity. The binding of a 14-mer oligonucleotide to the hairpin rescued the self-cleavage in a concentration-dependent manner. This modified ribozyme was inserted into the 5' UTR of a reporter gene, and the resulting construct was used to demonstrate that it is possible to modulate the ribozyme activity in cellulo with the oligonucleotide. Subsequently, ribozymes possessing specialized aptamers respecting other logic gates were also successfully designed and found to be functional in vitro. To our knowledge, this is the first example of HDV ribozyme regulation by oligonucleotides, as well as the first allosteric regulation of HDV ribozyme in mammalian cells.
authors.affiliation: RNA group/Groupe ARN, Département de biochimie, Faculté de médecine et des sciences de la santé, Pavillon de Recherche Appliquée sur le Cancer, Université de Sherbrooke, 3201 Jean-Mignault, Sherbrooke, Québec J1E 4K8, Canada.
authors.forename: Samuel G
authors.initials: SG
authors.lastname: Rouleau
authors.forename: Rachel
authors.initials: R
authors.lastname: Jodoin
authors.forename: Martin
authors.initials: M
authors.lastname: Bisaillon
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Perreault
mesh_terms: D000494:Allosteric Regulation; D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D017930:Genes, Reporter; D057809:HEK293 Cells; D003698:Hepatitis Delta Virus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D052157:Aptamers, Nucleotide; D016337:RNA, Catalytic