Detection of biomolecules
AssigneeIntelligene Ltd.
InventorsNathan Asher, Yaron Tikochinski, Guido Krupp, Jacob Grinberg, Adam Friedmann
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Abstract
The present invention concerns a method for detecting the presence of a catalytically active ribozyme in a medium. The detection of the catalytically active ribozyme may be a goal by itself, or the ribozyme may serve as a reporter for the presence of other biomolecules in an assayed sample. The detection is carried out in a catalytic system wherein the presence of the active ribozyme serves to produce other active ribozymes in a positive-feedback amplificatory manner.
Claims29
A method for detecting the presence of a catalytically active initiation ribozyme in a medium, comprising the steps of:
- (a) providing a catalytic system comprising catalytic system ribozymes which are catalytically inactive or spatially confined such that they cannot exert their catalytic activity on their target, the target of the catalytic system ribozymes being other catalytic system ribozymes,
- wherein the catalytic activity of the catalytic system ribozymes on said other catalytic system ribozymes cause either:
- (i) activation of inactive catalytic system ribozymes, or
- (ii) release of spatially confined catalytic system ribozymes to allow them to reach their targets,
- wherein at least some of the catalytic system ribozymes are a target of the catalytic activity of the initiation ribozyme, the catalytic activity of the initiation ribozyme on said some of the catalytic system ribozymes being that of (i) or (ii) above, and
- wherein the catalytic activity of the catalytic system ribozymes causes a change in the detectable properties of a detectable label;
- (b) contacting the medium with said catalytic system;
- (c) providing conditions permitting said catalytically active initiation ribozyme and catalytically active catalytic system ribozymes to exert their catalytic activity, whereby the presence of a catalytically active initiation ribozyme gives rise to a reaction cascade in which catalytic system ribozymes are activated or freed into the medium;
- and
- (d) detecting said detectable properties, a change in said properties being an indication of the presence of an active initiation ribozyme in said medium.
A kit for detecting the presence of a catalytically active initiation ribozyme in a medium comprising:
- a catalytic system comprising
- (a) assay ribozymes which are catalytically inactive or spatially confined such that they cannot exert their catalytic activity on their target, the target of the assay ribozymes being other ribozymes of the catalytic system, their catalytic activity on such other ribozymes causing either:
- (i) activation of inactive assay ribozymes, on
- (ii) release of spatially confined assay ribozymes to allow them to reach their targets,
- at least some of the assay ribozymes of the catalytic system being a target of the catalytic activity of the initiation ribozyme, the catalytic activity of the initiation ribozyme on said some of the assay ribozymes being that of (i) or (ii) above;
- and
- (b) a detectable label having detectable properties, wherein the catalytic activity of the assay ribozymes cause a change in the detectable properties.
Dependencies are inferred from the claim text.