US 2003/0152992 A1Published application

Screen employing fluorescence anisotropy to identify compounds with affinity for nucleic acids

OwnerLance G. Laing, Jaime E. Arenas +3inventors

InventorsLance G. Laing, Jaime E. Arenas, Sharon T. Cload, James W. Lillie, Andrew A. Pakula

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Abstract

The present invention provides methods for screening for bioactive compounds, in particular those that bind to RNA sequences involved in the pathogenesis of disease or in regulation of a physiological function. The methods involve assessing the stability and/or the conformation of an RNA target in the presence and absence of test ligands, and identifying as a ligand any test ligand that causes a measurable change in target RNA stability and/or conformation. In a preferred embodiment, the effect of a ligand on target RNA stability and/or conformation is assessed by measuring the fluorescence polarization of a fluorescently labeled probe.

Claims9

Dependent claims

A method for identifying a ligand that binds a predetermined target RNA sequence, which comprises:

(a) selecting as test ligands a plurality of compounds not known to bind to the target RNA sequence;
(b) incubating the target RNA sequence with one or more fluorescently labeled probes in the presence of a test ligand to produce a test combination;
(c) incubating the target RNA sequence with said one or more probes in the absence of a test ligand to produce a control combination;
(d) measuring one or more fluorescence properties of said probe;
and
(e) selecting as a ligand any test ligand that causes a measurable change in at least one of the fluorescence properties measured in step (d) in the test combination relative to the control combination, wherein said property reflects the stability and/or conformation of said target RNA.

3 more elements

Dependencies are inferred from the claim text.

Facts

Application: 10/370,819Filed 20 Feb 2003
Priority: 5 Mar 1997
Published: 14 Aug 2003
Term ends: Not estimatedThe source records don’t support an estimate.

Classification10 codes

C40B 30/10
Methods of screening libraries: by measuring physical properties, e.g. mass
C12N 15/1048
Isolating an individual clone by screening libraries: selex
C12Q 1/6811
Involving nucleic acids: selection methods for production or design of target specific oligonucleotides or binding molecules
C12Q 1/6813
Involving nucleic acids: hybridisation assays

Family

Application60/039,911Us provisional application · filed 5 Mar 1997
Application09/060,449Continuation · filed 5 Mar 1998
Application09/961,008Continuation · filed 20 Sep 2001
Application10/370,819Continuation · filed 20 Feb 2003
US 2002/0031778 A1Published application
US 6,331,392 B1Related patent
US 6,569,628 B2Related patent
US 2003/0152992 A1This patent

From the US filing history. Foreign family members aren’t listed.