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US 6,670,193 B2Granted
Fluorescent nucleoside analogs and combinatorial fluorophore arrays comprising same
AssigneeResearch Corporation Technologies, Inc.
InventorEric T. Kool
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AssigneeResearch Corporation Technologies, Inc.
InventorEric T. Kool
Estimating the term…
The present invention provides fluorescent nucleoside analogs which comprise a fluorescent cyclic compound joined to a carbon of a sugar molecule such as pentose, hexose, ribose or deoxyribose or analogs thereof in either an α or β configuration. The subject compounds are useful as probes in the study of the structure and dynamics of nucleic acids and their complexes with proteins. In addition, the subject compounds are useful in any technique which uses labeled oligonucleotides for detection. Non-fluorescent spacer molecules in which a cyclohexane, cyclohexene, decalin, or benzene is joined to a carbon of a sugar moiety such as pentose, hexose, ribose or deoxyribose are also provided. Also provided are the 5′ dimethoxytrityl-3′-O-phosphoramidite derivatives, suitable for incorporation into oligonucleotides by automated synthesizers. Combinatorial fluorophore array (CFA) libraries comprising oligomers of the subject nucleoside analogs attached to one or more solid supports are also provided as are methods of selecting fluorophores from the CFA libraries. The present invention also provides oligonucleotide analogs comprising one or more of the subject nucleoside analogs in place of a DNA or RNA base.
A method of selecting a fluorophore suitable for use in labeling a nucleic acid molecule which comprises:
A method of identifying a fluorophore emitting large Stokes shifts which comprises (a) constructing a combinatorial fluorophore array library which comprises multiple solid supports or multiple locations on a solid support, wherein each support or location has attached thereto an oligomer comprising a nucleoside analog, said nucleoside analog being a fluorescent cyclic compound joined to a carbon at the C-1 position thereof of a sugar moiety in either an α or β configuration and wherein said sugar moiety is one of pentose or hexose, (b) exciting the library at short wavelengths, and (c) selecting a fluorophore which emits light at a much longer wavelength.
A method of identifying a fluorophore involved in energy transfer which comprises (a) constructing a combinatorial fluorophore array library (CFA) which comprises multiple solid supports or multiple locations on a solid support, wherein each support location has attached thereto an oligomer comprising a nucleoside analog, aid nucleoside analog being a fluorescent cyclic compound joined to a carbon at the C1 position of a sugar moiety in either an α or β configuration wherein said sugar moiety is a pentose or hexose, and one or more non-fluorescent nucleoside analogs selected from the group consisting of cyclohexene-2 deoxyriboside, cyclohexene-2-deoxyribose, declaim 2-deoxyriboside, and benzene-2 deoxyriboside wherein said one or more non-florescent nucleoside analogs is interspaced between the fluorescent nucleoside analogs;
The method of identifying a fluorophore involved in energy transfer which comprises:
A method for identifying a fluorophore sequence that changes its emission wavelength or intensity on binding an analyte, which method comprises:
Dependencies are inferred from the claim text.