title: Trans-cleaving hammerhead ribozymes with tertiary stabilizing motifs: in vitro and in vivo activity against a structured viroid RNA.
abstract: Trans-cleaving hammerheads with discontinuous or extended stem I and with tertiary stabilizing motifs (TSMs) have been tested previously against short RNA substrates in vitro at low Mg(2+) concentration. However, the potential of these ribozymes for targeting longer and structured RNAs in vitro and in vivo has not been examined. Here, we report the in vitro cleavage of short RNAs and of a 464-nt highly structured RNA from potato spindle tuber viroid (PSTVd) by hammerheads with discontinuous and extended formats at submillimolar Mg(2+). Under these conditions, hammerheads derived from eggplant latent viroid and peach latent mosaic viroid (PLMVd) with discontinuous and extended formats, respectively, where the most active. Furthermore, a PLMVd-derived hammerhead with natural TSMs showed activity in vivo against the same long substrate and interfered with systemic PSTVd infection, thus reinforcing the idea that this class of ribozymes has potential to control pathogenic RNA replicons.
authors.affiliation: Instituto de Biología Molecular y Celular de Plantas, UPV-CSIC, Campus Universidad Politécnica de Valencia, Avenida de los Naranjos, 46022 Valencia, Spain.
authors.forename: Alberto
authors.initials: A
authors.lastname: Carbonell
authors.forename: Ricardo
authors.initials: R
authors.lastname: Flores
authors.forename: Selma
authors.initials: S
authors.lastname: Gago
mesh_terms: D001483:Base Sequence; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*; D014772:Viroids / Q000235:genetics*
chemical_list: D016337:RNA, Catalytic; D012367:RNA, Viral; C427204:hammerhead ribozyme; D012313:RNA