Eastman-kodak Company

Sterling Drug Inc.

Cetus Corporation

Sanofi

Johnson & Johnson Clinical Diagnostics, Inc.

Fred T Oakes

Harold C Warren

Frederick H Hausheer

John Bruce Findlay

Alexander L Weis

Corey H Levenson

Chu-an Chang

Paul F Cavanaugh, Jr

Patricia S Moskwa

Ignazio S Ponticello

Richard C Sutton

Susan J Danielson

Bruce E Babb

Marsha D Oenick

Prasad V Chaturvedula

Thomas J Cummins

Brent A Burdick

Daniel J Delecki

Marlene M King

Nancy F Green

Krishna Jayaraman

Janice M Mayer

The present invention relates to compounds, compositions and methods for inhibiting gene expression. The compounds of this invention comprise 1) oligonucleoside sequences of from about 6 to about 200 bases having a three atom internucleoside linkage or 2) oligonucleotide sequences of from about 9 to about 200 bases having a diol at either or both termini.

Oligonucleotide analogues containing phosphate diester linkage

Compounds, compositions and methods for inhibiting gene expression are disclosed. The compounds comprise oligonucleotide sequences of from about 9 to about 200 bases having a diol at either or both termini. Preferred diols are polyalkyleneglycols, preferably polyethyleneglycols. Pharmaceutical compositions comprising the compounds and a physiologically acceptable carrier and methods of inhibiting gene expression in mammals comprising administering such compounds are also provided. Methods for inhibiting nuclease cleavage of compounds are also provided.

Nuclease resistant compounds

A nucleic acid test article can be used to detect a targeted nucleic acid found in a specimen. The test article includes a substrate having two opposing surfaces and a water-insoluble nucleic acid probe attached in a distinct zone of one of the surfaces. The probe is prepared from a water-insoluble particle to which is covalently attached an oligonucleotide which is complementary to the targeted nucleic acid. Substantially none of the probe is embedded within the surface of the substrate. Particularly useful test articles have a multiplicity of water-insoluble probes located in distinct zones on one of the substrate surfaces. These probes are useful for the detection of a multiplicity of targeted nucleic acids, particularly after amplification by polymerase chain reaction.

Nucleic acid test article and its use to detect a predetermined nucleic

Amplification, replication or detection of a predetermined target nucleic acid can be carried out using a unique primer composition. This composition comprises and aqueous mixture of a first oligonucleotide primer which is substantially complementary to a first nucleic acid sequence of the target, but which is suspected of having one or more mismatches with the target at or near its 3' end. Also included in the composition is one or more additional primers which are complementary to a nucleic acid sequence of the target. This sequence is either: (i) inclusive of only a portion of the first nucleic acid sequence, (ii) immediately adjacent to the first nucleic acid sequence, or (iii) removed from the first sequence by one or more bases, but which additional primer is capable of forming a primer extension product complementary to the first sequence. These composition components can be supplied as part of a diagnostic test kit which can include other regents if desired.

Diagnostic kit, primer composition and their use for replication or

There are described a device and method for doing confined reactions such as PCR amplification and detection, wherein solids (e.g., beads) used to obtain separation between bound and 'free' label reagents, are transferred from region to region, specifically through a wash liquid so as to wash off the 'free' label reagent from the solids. Separate chambers have dividers that are overcome by piercing or by liquification, to create passageways for the transfer of the solids. The passageways remain contained within the device.

Device for moving a target-bearing solid through a liquid for detection

A method is provided for preparing a covalent conjugate of an oligonucleotide and an enzyme, such as peroxidase. This conjugate can be used as a probe in hybridization assays and in polymerase chain reaction procedures. The method generally comprises the steps of: reacting an enzyme having a reactive amino group with a mercapto-substituted organic compound to form a blocked intermediate, removing the blocking group to form an enzyme reagent, and reacting the enzyme reagent with a functionalized oligonucleotide reagent.

Method for preparing a covalent conjugate of an oligonucleotide and an

Specific binding methods are used for diagnostic assays and purification separations whereby the specific binding capture reagent is prepared from copolymers having highly reactive carboxy groups. These groups are extended from the polymer surface with a linking group having from 8 to 50 atoms in the chain and two or more alkylene, arylene, alkylenearylene or arylenealkylene groups. To these reactive groups is attached a biologically active substance such as a protein or oligonucleotide which then participates in the diagnostic assays or purification separation methods.

Specific binding analytical and separation methods using carboxy

A covalent conjugate of an enzyme, such as peroxidase, glucose oxidase, alkaline phosphatase and beta-galactosidase, and an oligonucleotide is herein disclosed. This conjugate can be used as a probe in hybridization assays and in polymerase chain reaction procedures.

Oligonucleotide-enzyme conjugate that can be used as a probe in

Exonuclease resistant terminally substituted oligonucleotides

Certain phenalenimine fluorescent compounds represented by the structure ##STR1## wherein R' and R' are independently hydrogen, alkyl, cycloalkyl, aryl or a heterocycle, or R' comprises the carbon and heteroatoms which form a fused ring with the compound nucleus, are useful in biomedical and analytical determinations. These compounds can be used for staining cells, as well as for the determination of various analytes found in human or animal biological fluids. Such determinations can be carried out in solution or by using dry analytical elements. The fluorescent compounds can be reacted with quinone nuclei to form reducible compounds which are also useful in analytical methods. In addition, the compounds can be incorporated into what are known as 'loadable' latex particles to form detectable labels and biological reagents.

Phenalenimine fluorescent dyes and their use in analytical compositions,

Biologically active reagents are prepared from particles of copolymers having highly reactive carboxy or equivalent groups. The reagents are prepared by covalently attaching biologically active substances, for example antibodies, to the particles, directly or indirectly through highly reactive carboxy groups on the particle surface. These reagents are used to advantage in analytical elements, methods for the detection of specific binding ligands (such as immunological species) and immunoassays, and in purification methods as affinity chromatography reagents.

Biologically active reagents prepared from carboxy-containing polymer,

A method of synthesizing double-stranded DNA sequences is disclosed. The method comprises the steps of:

Method of making double-stranded DNA sequences

Certain phenalenimine fluorescent compounds represented by the structure ##STR1## wherein R' and R' are independently hydrogen, alkyl, cycloalkyl, aryl or a heterocycle, or R' comprises the carbon and heteroatoms which forms a fused ring with the compound nucleus, are useful in biomedical and analytical determinations. These compounds can be used for staining cells, as well as for the determination of various analytes found in human or animal biological fluids. Such determination can be carried out in solution or by using dry analytical elements. The fluorescent compounds can be reacted with quinone nuclei to form reducible compounds which are also useful in analytical methods. In addition, the compounds can be incorporated into what are known as 'loadable' latex particles to form detectable labels and biological reagents.

Phenalenimine Fluorescent dyes and their use in analytical methods

Oligonucleotides derivatized to incorporate horseradish peroxidase (HRP) are useful in diagnostic methods and provide significant advantage over radioactively labeled oligonucleotide probes. The HRP-derivatized oligonucleotides can be easily and efficiently prepared by reacting a sulfhydryl containing oligonucleotide with a conjugate of a mal-sac-HNSA ester and HRP under mild and easily controlled reaction conditions.

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Rochester, NY, United States of America

Fred T Oakes