The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Feb. 25, 2025

Filed:

Oct. 01, 2024
Applicants:

Fenfei Leng, Palmetto Bay, FL (US);

Matthew Dias, Miami, FL (US);

Inventors:

Fenfei Leng, Palmetto Bay, FL (US);

Matthew Dias, Miami, FL (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12Q 1/682 (2018.01); C12Q 1/37 (2006.01); C12Q 1/44 (2006.01); C12Q 1/533 (2006.01); C12Q 1/6832 (2018.01);
U.S. Cl.
CPC ...
C12Q 1/682 (2013.01); C12Q 1/37 (2013.01); C12Q 1/44 (2013.01); C12Q 1/533 (2013.01); C12Q 1/6832 (2013.01);
Abstract

The subject invention provides a novel fluorescence-based, T5 exonuclease-amplified DNA cleavage assay for gyrase poisoning inhibitor discovery. According to the assay, multiple gyrase molecules can simultaneously bind to a plasmid DNA molecule to form multiple gyrase-DNA cleavage complexes on the same plasmid. These gyrase-DNA cleavage complexes, greatly stabilized by a gyrase poisoning inhibitor, and can be trapped by a detergent such as sarkosyl. Digestion of gyrase by proteinase K results in the production of small DNA fragments, which can be digested by T5 exonuclease. This fluorescence-based DNA cleavage HTS assay is also suitable for screening large compound libraries to identify inhibitors against DNA topoisomerases, including human DNA topoisomerase IIα.


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