Abstract
DNA
topoisomerases (Topos) are enzymes responsible for the relaxation of the
positive and negative supercoiled DNA as well as the resolution of DNA knots
and catenanes during vital biological processes. DNA Topos create transient DNA
break which results in changes in DNA topology, a DNA property that affects
genomic stability and the susceptibility to cancers or certain hereditary
diseases. Therefore, DNA Topos are an important target for certain antibiotics
and/or anticancer drugs.Commonly
used assays to test DNA Topos activities include agarose gel electrophoresis
and fluorescently labeled DNA molecules. However, the agarose gel
electrophoresis method is labor-intensive, time-consuming, and does not apply
to high throughput screening (HTS) assays. On the other hand, using
fluorescently labeled DNA molecules is expensive and the method could interfere
with the final detection signal. Hence, there is a need for methods and assays to
identify Topo inhibitors as potential initiators for drug development. FIU
scientists have developed assays and to screen compound libraries to identify
potential inhibitors targeting different DNA topoisomerases, such as bacterial
DNA topoisomerases I and II, and DNA gyrases. The method is based on using a
circular plasmid DNA, with at least one hairpin structure, and T5 exonuclease
(T5E) ability to digest the circular plasmid DNA in a specific configuration.
Benefit
Applicable for high throughput screening assaysCost-effective assaysSensitive assays
Market Application
Drug development
Brochure
