Abstract: Electrochemiluminescent enzymes, their preparation and use as biosensors are disclosed. Specifically, two appendages are covalently attached to a desired dehydrogenase enzyme; (1) a nicotinamide adenine cofactor or analog thereof, and (2) a luminescent ruthenium complex. For example, glucose concentrations is the following way. A doubly-modified glucose dehydrogenase could oxidize glucose with concomitant reduction of the attached NAD+ to NADH. Because NADH, but not NAD+, is able to interact with surface ruthenium to promote ECL, only enzyme molecules that have reacted with glucose will emit light from their ruthenium label in an ECL instrument. The relative close proximity of NADH and ruthenium on the enzyme surface enhances light emission as compared to the same concentrations in free solution. When NADH reduces ruthenium, it returns to become NAD+, permitting multiple cycles of ECL light emission from a single enzyme molecule. Such biosensors can be used in solution or bound to a solid surface.

Abstract: A method of selecting for missense mutants that express a protein is disclosed. The method comprises constructing an operon comprising an upstream gene and a downstream reporter gene, wherein the upstream gene and the reporter gene are translationally coupled, wherein the reporter protein is expressed after the translation of the upstream gene. The method further comprises expressing the downstream reporter gene after introducing at least one mutation into the upstream gene. Finally, the mutants are screened for the ability to express the reporter protein, which is indicative of a missense mutant.

Abstract: Nucleic acid molecule which modulates the synthesis, expression and/or stability of an mRNA encoding one or more receptors of vascular endothelial growth factor.

Abstract: The present invention relates to the discovery in eukaryotic cells, particularly mammalian cells, of a novel family of cell-cycle regulatory proteins (“CCR-proteins”). As described herein, this family of proteins is characterized by four ankyrin repeats and the ability to bind to a cyclin dependent kinase (CDK). The family includes a polypeptide having an apparent molecular weight of 16 kDa, and a polypeptide having an apparent molecular weight of approximately 15 kDa, each of which can function as an inhibitor of cell-cycle progression, and therefore ultimately of cell growth. Thus, similar to the role of p21 to the p53 checkpoint, the subject CCR-proteins may function coordinately with the cell-cycle regulatory protein, retinoblastoma (RB).

Abstract: In accordance with the present invention, antisense oligonucleotides are provided with enhanced membrane permeability and stability. This is accomplished in accordance with the invention through conjugating oligoribonucleotides with a hydrophobic carrier agent at the 2′-O position of the oligonucleotides. The hydrophobic carrier agent comprises a compound of the following general structure: wherein R1, R2, R3, R4, and R5 are independently H, NO2, halide, linear or branched alkyl, linear or branched acyl, linear or branched alkylene, linear or branched O-alkyl, linear or branched amido, linear or branched S-alkyl, mono or disubstituted amine, linear or branched thioamido, phosphothionate, or phosphothioate. In a preferred embodiment, R1 and R3 are NO2. In such embodiment, it will be appreciated that when R2, R4, and R5 are H, the compound is DNP and when R4 is F, the compound is FDNP.

Abstract: The present invention relates to ubiquitin-specific thiol proteases or deubiquitinating enzymes, referred to as DUB (DeUBiquitinating) enzymes, of eukaryotic origin which are members of a superfamily of deubiquitinating enzymes and which comprise a new subfamily of deubiquitinating enzymes which are similar in size and amino acid sequence to one another. DUB enzymes of the present invention are of eukaryotic origin, such as vertebrate origin, including mammalian (e.g., murine, human) origin, as well as yeast origin. All DUB enzymes of the present invention are inducible by at least one cytokine.

Abstract: A method of inhibiting the proliferation and causing the differentiation of undifferentiated cells comprising contacting the undifferentiated cells with an effective amount of an antisense oligonucleotide having a sequence which is complementary to a region of the IGF-1 receptor RNA. The sequence of the antisense oligonucleotide is selected from an oligodeoxynucleotide sequence complementary to codons −29 to −24 of the signal sequence of the IGF-1 receptor and an oligoribodeoxynucleotide sequence complementary to codons 1 to 309 of the sequence of the IGF-1 receptor. The oligoribodeoxynucleotide sequence may be provided by an expression vector.

Abstract: The invention includes novel fusion nucleic acids encoding fusion polypeptides which when expressed in a filamentous fungus result in the expression of fusion polypeptides. The fusion nucleic acids comprise four nucleic acids which encode a fusion polypeptide comprising first, second, third and fourth amino acid sequences. The first nucleic acid encodes a signal polypeptide functional as a secretory sequence in a first filamentous fungus. The second nucleic acid encodes a secreted polypeptide or functional portion thereof which is normally secreted from the same filamentous fungus or a second filamentous fungus. The third nucleic acid encodes a cleavable linker while the fourth nucleic acid comprises at least two nucleic acids encoding desired polypeptides.

Abstract: The present invention relates to methods and compositions for the elucidation of mammalian gene function. Specifically, the present invention relates to methods and compositions for improved mammalian complementation screening, functional inactivation of specific essential or non-essential mammalian genes, and identification of mammalian genes which are modulated in response to specific stimuli. In particular, the compositions of the present invention include, but are not limited to, replication-deficient retroviral vectors, libraries comprising such vectors, retroviral particles produced by such vectors in conjunction with retroviral packaging cell lines, integrated provirus sequences derived from the retroviral particles of the invention and circularized provirus sequences which have been excised from the integrated provirus sequences of the invention. The compositions of the present invention further include novel retroviral packaging cell lines.

Abstract: This invention provides a method of determining a chromosomal breakpoint in a subject suffering from multiple myeloma which comprises steps of: (a) obtaining a DNA sample from the subject suffering from multiple myeloma; (b) determining whether there is J and C disjunction in the immunoglobulin heavy chain gene in the obtained DNA sample; (c) obtaining a genomic library having clones which contain genomic DNA fragments from the DNA sample which shows positive J and C disjunction; (d) selecting and isolating clones of the obtained library which show positive hybridization with a probe which is capable of specifically hybridizing with the C but not the J region of the immunoglobulin heavy chain gene; (e) preparing fluorescent probes from the genomic DNA fragments of the isolated clones from step (d); (f) hybridizing said fluorescent probes with metaphase chromosomes; and (g) determining the identity of the chromosomes which are capable of hybridizing to said fluorescent probes, wherein the identification of a c

Abstract: The invention provides novel tumor suppressor genes, methods for making and using these and related tumor suppressor genes and proteins and peptides, and nucleic acids encoding these and related tumor suppressor proteins and peptides.

Abstract: Compositions and methods are provided for modulating the expression of PKC-&dgr; and for the treatment and diagnosis of diseases associated with protein kinase C-&dgr;. Methods of treating animals suffering from disease amenable to therapeutic intervention by modulating protein kinase C-&dgr; expression with an oligonucleotide specifically hybridizable with RNA or DNA corresponding to PKC-&dgr; are disclosed. Methods of modulating the expression of TNF-a using the compositions of the present invention are also provided.

Abstract: The current invention concerns the use of a receptor from the RZR/ROR receptor family or of a functional fragment thereof in a test of a compound for anti-autoimmune, anti-arthritic, anti-tumor, melatonin-like and/or melatonin-antagonistic activity and the production of a receptor ligand complex comprising said receptor or a functional fragment thereof and a ligand of said receptor. Described is also a method for testing compounds for said activity (screening for ligands) and the active compounds identified therewith.

Abstract: The present invention is based on the finding that nucleic acids containing at least one unmethylated cytosine-guanine (CpG) dinucleotide affect immune responses in a subject. These nucleic acids containing at least one unmethylated cytosine-guanine (CpG) dinucleotide can be used to treat pulmonary disorders having an immunologic component, such as a response to inhaled lipopolysaccharide. The invention provides methods of treating subjects who have or are at risk of having these pulmonary disorders, and methods of altering the immunological components of the pulmonary disorders. The invention also provides pharmaceutical compositions for treating pulmonary disorders that have an immunologic component.

Abstract: The present invention provides methods for increasing ribozyme catalytic activity without reducing specificity, which methods comprise contacting an RNA molecule with a ribozyme having a flanking sequence modified to contain 2′-O-substituted nucleotides. The invention also provides ribozymes comprising a flanking sequence modified to contain 2′-O-substituted nucleotides. In addition, the invention provides methods for increasing ribozyme catalytic activity comprising contacting an RNA molecule with a ribozyme having a flanking sequence modified to contain a 2′-O-substituted nucleotide and a facilitator oligonucleotide. The present invention further provides compositions comprising a ribozyme having modified flanking sequences and an effective amount of a facilitator oligonucleotide.

Abstract: Retroviral vectors contain cis-acting viral elements for the expression, encapsidation, reverse transcription and integration of the retroviral genome nucleic acid sequence. However, these elements are not useful in the integrated provirus and may cause many problems. A retroviral vector is provided for eliminating most of the viral elements. This vector uses, among other things, the bacteriophage P1 Cre-lox recombination system. The 32-nucleotide loxP site is inserted into 3′LTR sequence U3 with the gene to be inserted into the cell. After loxP duplication using the LTR, the LTRs may be recombined by enzyme Cre. The structure of the resulting provirus in the host genome corresponds to a single LTR carrying a single copy of the gene to be inserted into the cell. If the Cre expression unit is inserted between the two LTRS, only single-LTR proviral structures are found following infection with the retroviral vector.

Abstract: Antisense compounds, compositions and methods are provided for modulating the expression of SHP-2. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding SHP-2. Methods of using these compounds for modulation of SHP-2 expression and for treatment of diseases associated with expression of SHP-2 are provided.

Abstract: A method for increasing the specificity of hybridization between a nucleic acid probe and a nucleic acid sequence to be detected, by addition of blocker molecules, which are complementary to the probe, raise of temperature in order to melt non-perfectly matched hybrids of probe and detected nucleic acid sequences, and lowering of the temperature again.

Abstract: The promoter of the EPCR gene has been isolated from both murine (SEQ. ID No. 1) and human (SEQ. ID No. 2) genomic libraries. The promoter includes a region (nucleotides 3130 to 3350 of SEQ. ID No. 1 which affects selective gene expression in endothelial cells), and a region (nucleotides 2270 to 2840 of SEQ. ID No. 1) which affects selective gene expression in large vessel endothelial cells, as compared to expression in all endothelial cells. The EPCR promoter contains a thrombin responsive element, CCCACCCC (SEQ. ID No. 3), (murine, nucleotides 3007 to 3014 SEQ. ID No. 1 and human, nucleotides 2722 to 2729 SEQ. ID No. 2). The EPCR also contains a serum response element (nucleotides 2990 to 3061 of SEQ. ID No. 1). The regulatory sequences present in the EPCR promoter can be used for thrombin or serum controlled recombinant gene expression specific to either all endothelial cells or primarily endothelial cells of large vessels.

Abstract: The present invention relates to a method of detecting a DNA sequence by means of a DNA:DNA hybrid in real time using fluorescence. The present invention eliminates the need to use radioactive probes to detect the DNA and eliminates the delay needed for autoradiographic exposure of the X-ray to the radioactive label.