Kristal Domenico - Academia.edu (original) (raw)

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Papers by Kristal Domenico

Research paper thumbnail of Selection, design, and characterization of a new potentially therapeutic ribozyme

RNA, 2002

An in vitro selection was designed to identify RNA-cleaving ribozymes predisposed for function as... more An in vitro selection was designed to identify RNA-cleaving ribozymes predisposed for function as a drug. The selection scheme required the catalyst to be transacting with phosphodiesterase activity targeting a fragment of the Kras mRNA under simulated physiological conditions. To increase stabilization against nucleases and to offer the potential for improved functionality, modified sequence space was sampled by transcribing with the following NTPs: 29-F-ATP, 29-F-UTP, or 29-F-5-[(N-imidazole-4-acetyl) propylamine]-UTP, 29-NH 2-CTP, and GTP. Active motifs were identified and assessed for their modified NMP and divalent metal dependence. The minimization of the ribozyme's size and the ability to substitute 29-OMe for 29-F and 29-NH 2 moieties yielded the motif from these selections most suited for both nuclease stability and therapeutic development. This motif requires only two 29-NH 2-Cs and functions as a 36-mer. Its substrate sequence requirements were determined to be 59-Y-G-H-39. Its half-life in human serum is .100 h. In physiologically relevant magnesium concentrations [;1 mM] its k cat 5 0.07 min-1 , K m 5 70 nM. This report presents a novel nuclease stable ribozyme, designated Zinzyme TM , possessing optimal activity in simulated physiological conditions and ready for testing in a therapeutic setting.

Research paper thumbnail of Selection, design, and characterization of a new potentially therapeutic ribozyme

RNA, 2002

An in vitro selection was designed to identify RNA-cleaving ribozymes predisposed for function as... more An in vitro selection was designed to identify RNA-cleaving ribozymes predisposed for function as a drug. The selection scheme required the catalyst to be transacting with phosphodiesterase activity targeting a fragment of the Kras mRNA under simulated physiological conditions. To increase stabilization against nucleases and to offer the potential for improved functionality, modified sequence space was sampled by transcribing with the following NTPs: 29-F-ATP, 29-F-UTP, or 29-F-5-[(N-imidazole-4-acetyl) propylamine]-UTP, 29-NH 2-CTP, and GTP. Active motifs were identified and assessed for their modified NMP and divalent metal dependence. The minimization of the ribozyme's size and the ability to substitute 29-OMe for 29-F and 29-NH 2 moieties yielded the motif from these selections most suited for both nuclease stability and therapeutic development. This motif requires only two 29-NH 2-Cs and functions as a 36-mer. Its substrate sequence requirements were determined to be 59-Y-G-H-39. Its half-life in human serum is .100 h. In physiologically relevant magnesium concentrations [;1 mM] its k cat 5 0.07 min-1 , K m 5 70 nM. This report presents a novel nuclease stable ribozyme, designated Zinzyme TM , possessing optimal activity in simulated physiological conditions and ready for testing in a therapeutic setting.

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