title: Hairpin ribozyme specificity in vivo: a case of promiscuous cleavage.
abstract: We have used differential display to address the question of ribozyme specificity in vivo. Stably transfected PC12 cells bearing either a hairpin ribozyme expression plasmid targeted to betaAPP mRNA or the vector alone were analyzed using nine different primer pairs. One of the few differentially expressed genes obtained from this screen corresponded to rat ribosomal protein L19. Steady-state levels of L19 mRNA were lower in ribozyme-transfected cells compared to either vector-transfected cells or native PC12 cells, and a sequence within the L19 message was cleaved by the betaAPP hairpin ribozyme in vitro. These data imply that sequence-specific unintended cleavage of non-target mRNAs may present a formidable problem to the use of hairpin ribozyme therapeutic agents.
authors: Laboratory of Molecular Neurobiology, Department of Molecular Biology, New York State Institute for Basic Research in Developmental Disabilities, 1050 Forest Hill Road, Staten Island, New York, 10314, USA. bob1028@interport.net
authors: R B
authors: RB
authors: Denman
mesh_terms: D016564:Amyloid beta-Protein Precursor / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015536:Down-Regulation; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D009690:Nucleic Acid Conformation; D016716:PC12 Cells; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics*; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity; D014162:Transfection
chemical_list: D016564:Amyloid beta-Protein Precursor; D016337:RNA, Catalytic; D012333:RNA, Messenger; C489092:RPL19 protein, human; D012269:Ribosomal Proteins; C050482:Rpl19 protein, rat