Methods and materials for large-scale assessment of ligand binding selectivity of g-quadruplex recognition using custom g4 microarrays
AssigneesPurdue Research Foundation, The United States of America, as represented by the Secretary, Department of Health and Human Servic
InventorsDanzhou YANG, Guanhui WU, Charles VINSON
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Abstract
Described herein are devices and processes using single-stranded DNA sequences capable of forming G-quadruplexes (G4s) to assess the binding affinity and binding selectivity of potential G4-interactive ligands.
Claims20
A method for determining binding preferences of a non-fluorescent test compound for one or more target G-quadruplex moieties, the method comprising;
- a) incubating a device comprising a plurality of single-stranded nucleic acid molecules capable of forming one or more G-quadruplex moieties including the target G-quadruplex moieties with a solution comprising a G-quadruplex stabilizing cation selected from the group consisting of Na+ and K+;
- b) incubating the device with a solution of a compound capable of providing a fluorescent signal (a fluorescent compound), wherein the fluorescent compound is capable of binding to the target G-quadruplex moieties;
- c) measuring a first fluorescent signal from the fluorescent compound bound to the device;
- d) removing the fluorescent compound from the device;
- e) contacting the device with a solution of the fluorescent compound and the test compound;
- f) measuring a second fluorescent signal from the fluorescent compound bound to the device;
- and
- g) using the first fluorescent signal and the second fluorescent signal to calculate the binding preferences of the test compound.
A method for determining the binding preference of a test compound capable of providing a fluorescent signal (a fluorescent test compound) for one or more target G-quadruplex moieties, the method comprising the steps of;
- a) incubating a device comprising a plurality of single-stranded nucleic acid molecules capable of forming one or more G-quadruplex moieties including the target G-quadruplex moieties with a solution comprising a G-quadruplex stabilizing cation selected from the group consisting of Na+ and K+,
- b) contacting the fluorescent test compound with the device;
- c) measuring a first fluorescent signal from the fluorescent test compound bound to the device;
- d) incubating the device with a solution of solution of Li+;
- e) contacting the fluorescent test compound with the device;
- f) measuring a second fluorescent signal from the fluorescent test compound bound to the device;
- g) using the first fluorescent signal and the second fluorescent signal to calculate the binding preference of the fluorescent test compound.
Dependencies are inferred from the claim text.