In vitro selection of adenine-dependent ribozyme against Tpl2/Cot oncogene.
Hairpin ribozymes possess the properties of RNA sequence-specific recognition and site-specific cleavage. These properties make them a powerful extension of the antisense approach for the inhibition of gene expression. From a randomized RNA pool of hairpin ribozymes, using the systematic evolution of ligands by exponential enrichment, we have obtained an adenine-dependent hairpin ribozyme, Tpl2/Cot (tumour progression locus 2) ribozyme, which cleaves the Tpl2/Cot kinase mRNA sequence at nucleotides A225/G226 relative to the start codon of translation. This serine/threonine kinase activates the mitogen-activated protein kinase pathway implicated in cell proliferation in cancer. The selected 'Tpl2/Cot-YL ribozyme' efficiently cleaves its target sequence in cis and in trans; furthermore, the ribozyme efficiently cleaves a longer target sequence of 54 nucleotides in trans, as well as the full-length mRNA.
Li
Yan-Li
YL
Institut Jacques-Monod, Laboratoire de Biochimie de l'Evolution et Adaptabilité Moléculaire, UPMC Université Paris 06, Paris, France.
Vergne
Jacques
J
Torchet
Claire
C
Maurel
Marie-Christine
MC
D000225:Adenine / Q000494:pharmacology*; D001483:Base Sequence; D017124:Conserved Sequence; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D020930:MAP Kinase Kinase Kinases / Q000187:drug effects / Q000235:genetics*; D015636:Magnesium Chloride / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011518:Proto-Oncogene Proteins / Q000187:drug effects / Q000235:genetics*; D011519:Proto-Oncogenes; D016337:RNA, Catalytic / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics*; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
D011518:Proto-Oncogene Proteins; D016337:RNA, Catalytic; D012333:RNA, Messenger; D015636:Magnesium Chloride; D020930:MAP Kinase Kinase Kinases; C069937:MAP3K8 protein, human; D000225:Adenine