High-throughput screening assay for inhibitors of nucleic acid polymerases

US5635349A · kind A · utility

Assignee

• Tularik Limited · US

Inventors

• Kelly Lamarco · San Francisco, US
• Berta Strulovici · San Francisco, US
• Pengguang Wu · San Francisco, US

Key dates

• Filing date: Dec 2, 1994
• Grant date: Jun 3, 1997
• Expiry date: Dec 2, 2014

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/68
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The invention provides promoter specific and promoter non-specific screening assays for identifying an inhibitor of a pathogenic nucleic acid polymerase activity, e.g. an RNA polymerase derived from a pathogenic infectious organism such as a bacterium, protozoan or fungus. Generally, the methods involve forming a mixture of nucleotides, a polynucleotide template, a pathogenic polymerase candidate inhibitor of polymerase activity, where at least one of the nucleotides comprises a detectable label. The mixture is incubated under conditions whereby, but for the presence of the candidate inhibitor, the polymerase transcribes the polynucleotide template by catalyzing the polymerization of the nucleoside triphosphates into a polynucleotide having a nucleotide sequence complementary to that of the polynucleotide template. The nascent transcript is captured by a polynucleotide-selective agent immobilized on a solid substrate for subsequent washing and label detection.