Assay for Ribozyme target site

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 912, 536 251, C12Q 168

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055254683

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.

REFERENCES:
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Usman et al. "Automated Chemical Synthesis of Long Oligoribonucleotides Using 2 '-O-Silylated Ribonucleoside 3 '-O-Phosphoramidites on a Controlled-Pore Glass Support: Synthesis of a 43-Nucleotide Sequence Similar to the 3'-Half Molecule of an Escherichia coil Formylmethioine tRNA", 109 Jrnl of Amer. Chem. Society, 7845 1987.
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Hampel et al., "RNA Catalyst for Cleaving Specific RNA Sequence", filed Sep. 20, 1989, which is a Continuation-in-Part of U. S. Serial No. 07/247,100 filed Sep. 20, 1988.
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