Assay for Ribozyme target site
AssigneeRibozyme Pharmaceuticals, Inc.
InventorJames A. McSwiggen
Life of this patent
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Abstract
Method for determining target site accessibility for a ribozyme by providing a DNA oligonucleotide having a nucleotide sequence complementary to the RNA target. The DNA oligonucleotide is contacted with the RNA target in the presence of an agent, such as RNAseH, such that the RNAseH cleaves the RNA target when the DNA and RNA form a duplex. The occurrence of cleavage is detected by any standard methodology. Those DNA oligonucleotides which provide a detectable level of cleavage indicate that the RNA target site is accessible to ribozyme.
Claims15
A method for determining ribozyme target site accessibility and synthesis of a ribozyme, comprising the step of:
- contacting a DNA oligonucleotide complementary to the target RNA with said target RNA in the presence of RNaseH under RNA cleaving conditions;
- wherein cleavage by RNaseH indicates accessibility of said target RNA;
- and synthesizing a ribozyme to a said accessible target RNA.
A method for identifying a region of RNA accessible to cleavage by a ribozyme, comprising the step of:
- contacting a plurality of DNA oligonucleotides complementary to the RNA to be cleaved in the presence of RNaseH adapted to cleave the RNA of an RNA-DNA duplex under RNA cleaving conditions, wherein said DNA oligonucleotides are selected to hybridize to the RNA at a plurality of regions;
- and detecting cleavage of the RNA as an indicator of the most accessible region of the RNA to a ribozyme.
Dependencies are inferred from the claim text.