A novel allosterically trans-activated ribozyme, the maxizyme, with exceptional specificity in vitro and in vivo.
We have constructed an allosterically controllable novel enzyme (designated maxizyme) that can be transcribed in vivo under the control of a human tRNA(Val) promoter. The maxizyme has sensor arms that can recognize target sequences, and in the presence of such a target sequence only, it can form a cavity that can capture catalytically indispensable Mg2+ ions. As a target for a demonstration of the potential utility of the maxizyme, we chose BCR-ABL mRNA, the translated products of which cause chronic myelogenous leukemia. Only the maxizyme (but not conventional ribozymes) had extremely high specificity and high-level activity, not only in vitro but also in cultured cells including BV173 cells derived from a patient with a Philadelphia chromosome. The maxizyme induced apoptosis only in leukemic cells with this chromosome.
Kuwabara
T
T
National Institute for Advanced Interdisciplinary Research, Tsukuba Science City, Japan.
Warashina
M
M
Tanabe
T
T
Tani
K
K
Asano
S
S
Taira
K
K
D000494:Allosteric Regulation; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D002461:Cell Line, Transformed; D003593:Cytoplasm / Q000378:metabolism; D019281:Dimerization; D004792:Enzyme Precursors / Q000378:metabolism; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D008274:Magnesium / Q000494:pharmacology; D051379:Mice; D009690:Nucleic Acid Conformation; D010677:Philadelphia Chromosome; D016337:RNA, Catalytic / Q000378:metabolism* / Q000627:therapeutic use; D012333:RNA, Messenger / Q000378:metabolism*; D012366:RNA, Transfer, Val / Q000235:genetics; D013379:Substrate Specificity; D014162:Transfection; D014407:Tumor Cells, Cultured
D004792:Enzyme Precursors; D007377:Interleukin-3; D016337:RNA, Catalytic; D012333:RNA, Messenger; D012366:RNA, Transfer, Val; D016044:Fusion Proteins, bcr-abl; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases; D008274:Magnesium