Induction of ribozyme activity by anti-ribozyme oligonucleotides

Komatsu, Yasuo; Yamashita, Shohei; Kazama, Nobuhiro; Ohtsuka, Eiko

doi:10.1093/nass/42.1.279

Cited by 3 publications

(3 citation statements)

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“…The high activity of the cis tsHHRz at low-Me 2+ concentration leads to cleavage during any exposure to Mg 2+ ions during the biochemical procedures required for labeling (kinasing or transcription). Although this technical limitation can be partially overcome by using antisense oligonucleotides (Kronenwett et al, 1996;Komatsu et al, 1999;Khvorova et al, 2003), the resulting ribozyme is cleaved to a signifi cant extent.…”

Section: Fluorescence-based Assay For Cis-ribozyme Activitymentioning

confidence: 99%

The Different Role of High-Affinity and Low-Affinity Metal Ions in Cleavage by a Tertiary Stabilized Cis Hammerhead Ribozyme from Tobacco Ringspot Virus

Kisseleva

Khvorova²,

Westhof

et al. 2008

Oligonucleotides

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Section: Fluorescence-based Assay For Cis-ribozyme Activitymentioning

confidence: 99%

The Different Role of High-Affinity and Low-Affinity Metal Ions in Cleavage by a Tertiary Stabilized Cis Hammerhead Ribozyme from Tobacco Ringspot Virus

Kisseleva

Khvorova²,

Westhof

et al. 2008

Oligonucleotides

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“…Many reports have described ribozymes whose activity is modulated by molecular recognition of an oligonucleotide (Lorsch and Szostak 1995;Porta and Lizardi 1995;Komatsu et al 1999;Robertson and Ellington 1999;Kuwabara et al 2000;Stojanovic et al 2001;Wang and Sen 2001;Burke et al 2002;Iyo et al 2002;Komatsu et al 2002), a protein (Robertson and Ellington 2001;Hartig et al 2002;Vaish et al 2002;Wang and Sen 2002), or a small molecule (Tang and Breaker 1997;Araki et al 1998;Koizumi et al 1999;Soukup and Breaker 1999;Robertson and Ellington 2000;Wang et al 2002a). In these systems, molecular recognition is directly converted into catalytic events that can be monitored in homogeneous solution phase assays (Hartig et al 2002;Vaish et al 2002;Mei et al 2003), ELISA-like assays (Seetharaman et al 2001), array-based assays (Hesselberth et al 2003), or by RT-PCR (Robertson and Ellington 1999).…”

Section: Introductionmentioning

confidence: 99%

“…The allosteric and "expanded" ribozymes reported to date promote substantial catalysis in the absence of target (Lorsch and Szostak 1995;Porta and Lizardi 1995;Komatsu et al 1999;Robertson and Ellington 1999;Kuwabara et al 2000;Stojanovic et al 2001;Wang and Sen 2001;Burke et al 2002;Iyo et al 2002;Komatsu et al 2002). Such targetindependent catalysis limits the ability of these ribozymes to detect nucleic acid targets present in substoichiometric amounts because low levels of a target result in only a marginal increase in observed rate.…”

Section: Introductionmentioning

confidence: 99%

Zeptomole detection of a viral nucleic acid using a target-activated ribozyme

Vaish¹,

Jadhav²,

Kossen³

et al. 2003

RNA

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We describe a strategy for the ultra-sensitive detection of nucleic acids using "half" ribozymes that are devoid of catalytic activity unless completed by a trans-acting target nucleic acid. The half-ribozyme concept was initially demonstrated using a construct derived from a multiple turnover Class I ligase. Iterative RNA selection was carried out to evolve this half-ribozyme into one activated by a conserved sequence present in the hepatitis C virus (HCV) genome. Following sequence optimization of substrate RNAs, this HCV-activated half-ribozyme displayed a maximal turnover rate of 69 min −1 (pH 8.3) and was induced in rate by approximately 2.6 × 10 9 -fold by the HCV target. It detected the HCV target oligonucleotide in the zeptomole range (6700 molecules), a sensitivity of detection roughly 2.6 × 10 6 -fold greater than that previously demonstrated by oligonucleotide-activated ribozymes, and one that is sufficient for molecular diagnostic applications.

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High-Throughput Ribozyme-Based Assays for Detection of Viral Nucleic Acids

Kossen

Vaish

Jadhav

et al. 2004

Chemistry & Biology

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Many reports have suggested that target-activated ribozymes hold potential value as detection reagents. We show that a "half"-ribozyme ligase is activated similarly by three unstructured oligoribonucleotides representing the major sequence variants of a hepatitis C virus 5'-untranslated region (5'-UTR) target and by a structured RNA corresponding to the entire 5'-UTR. Half-ribozyme ligation product was detected both in an ELISA-like assay and in an optical immunoassay through the use of hapten-carrying substrate RNAs. Both assay formats afford a limit of detection of approximately 1 x 10(6) HCV molecules (1.6 attomol, 330 fM), a sensitivity which compares favorably to that provided by standard immunoassays. These data suggest that target-activated ribozyme systems are a viable approach for the sensitive detection of viral nucleic acids using high-throughput platforms.

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Induction of ribozyme activity by anti-ribozyme oligonucleotides

Cited by 3 publications

References 0 publications

The Different Role of High-Affinity and Low-Affinity Metal Ions in Cleavage by a Tertiary Stabilized Cis Hammerhead Ribozyme from Tobacco Ringspot Virus

The Different Role of High-Affinity and Low-Affinity Metal Ions in Cleavage by a Tertiary Stabilized Cis Hammerhead Ribozyme from Tobacco Ringspot Virus

Zeptomole detection of a viral nucleic acid using a target-activated ribozyme

High-Throughput Ribozyme-Based Assays for Detection of Viral Nucleic Acids

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