Abstract: The present invention relates to transport proteins, in particular VP22 and homologues thereof, and to methods of delivering these proteins and any associated molecules to a target population of cells. This transport protein has applications in gene therapy and methods of targeting agents to cells where targeting oat high efficiency is required.

Abstract: Nucleic acid from which a polypeptide with utrophin function can be expressed, especially mini-genes and chimaeric constructs. Expression significantly decreases the severity of the dystrophic muscle phenotype in an animal model, indicating usefulness in treatment of muscular dystrophy. The nucleic acid and encoded polypeptides are also useful in screening for substances to modulate utrophin binding to actin and/or the dystrophin protein complex.

Abstract: A method is provided for inducing DNA synthesis in differentiated neurons. According to certain embodiments of the invention, a method for inducing DNA synthesis in a differentiated neuron is provided that includes obtaining a vector comprising nucleic acid encoding an E2F regulator and/or an E1A regulator, wherein the vector can be used to express the nucleic acid in a differentiated neuron, and transfecting a differentiated neuron with the vector.

Abstract: A complex is described that is deliverable to a cell comprising inserting a nucleic acid or other cargo into a reverse micelle. The reverse micelle has the property to compact the nucleic acid for easier delivery.

Abstract: A method for preparing a closed vesicle by a rehydration of a closed vesicle using a rehydration solution, wherein said closed vesicle comprises a dehydrated micelle particle or dehydrated amphipathic micelle bilayers, preferably a liposome loading a pharmaceutically active substance, and the method is characterized in that the rehydration is carried out at a low temperature such as in the range of from 0° C. to 10° C. The method can provides a rehydrated stable vesicle without a leak of the loaded substance.

Abstract: Photoreactivity in a cell is modulated by incorporating an isolated optical trigger on the surface or in the membrane of the cell. Exposure of a cell bearing incorporated optical triggers causes the generation of a measurable physiological signal.

Abstract: A complex that comprises (i) a nucleic acid, (ii) an integrin-binding component, for example, an intergrin-binding peptide, (iii) a polycationic nucleic acid-binding component, for example, oligolysine, and (iv) a lipid component, for example, a cationic liposome, has transfection activity.

Abstract: Antisense oligonucleotides that hybridize to segments of the pres1, S, C, and &egr; regions of the hepatitis B virus (HBV) RNA pregenome inhibit replication of the virus. Pharmaceutical compositions which contain these oligonucleotides as the active ingredients are effective against HBV infection.

Abstract: Antisense compounds, compositions and methods are provided for modulating the expression of Syntaxin 4 interacting protein. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding Syntaxin 4 interacting protein. Methods of using these compounds for modulation of Syntaxin 4 interacting protein expression and for treatment of diseases associated with expression of Syntaxin 4 interacting protein are provided.

Abstract: Liposomes have been shown to be very safe complexes with virtually no side effects. Utilizing liposomes for gene therapy has been hampered by their non-specific nature of gene delivery. This protein sequence contains a DNA binding region, an alpha helix region which will transverse the liposome wall, and a receptor binding region to allow for targeting to a specific cell type, in this case, a hepatocyte.

Abstract: The present invention relates to neurogenesis inducing genes. In particular, the present invention provides neurogenesis inducing genes coding for Zic proteins, vectors containing such genes, host cells containing such vectors, proteins produced by such host cells, antibodies raised to such proteins, and therapeutic agents or agents for gene therapy for nervous diseases.

Abstract: The present invention relates to a method of transfecting cells comprising applying cells directly onto nucleic acids which are immobilized in transfection complexes on a surface and which transfect the cells. Preferably, the nucleic acids are immobilized in an array. In another aspect of the present invention, the method further includes expression of the nucleic acids in the transfected cells. In yet another aspect of the present invention, the method further comprises detecting the expression of the nucleic acids in the transfected cells.

Abstract: Particle aggregation of lipid:nucleic acid complex particles is prevented by incorporating a non-cationic lipid into lipid:nucleic acid complex particles containing a cationic lipid and a nucleic acid polymer. The non-cationic lipid is a polyethylene glycol-based polymer.

Abstract: The present invention provides a chimeric protein which comprises a mutated DNA methyltransferase portion and a DNA binding protein portion that binds sufficiently close to a promoter sequence of a target gene which promoter sequence contains a methylation site, to specifically methylate the site and inhibit activity of the promoter and thus inhibit expression of the target gene. This invention also provides for a method for inhibiting the expression of a target gene which includes contacting a promoter of the target gene with the chimeric protein, so as to specifically methylate the promoter sequence of the target gene thus inhibiting expression of the target gene.

Abstract: The present invention provides for lipid:nucleic acid complexes that have increased shelf life and high transfection activity in vivo following intravenous injection, and methods of preparing such complexes. The methods generally involve contacting a nucleic acid with an organic polycation to produce a condensed nucleic acid, and then combining the condensed nucleic acid with a lipid comprising an amphiphilic cationic lipid to produce the lipid:nucleic acid complex. This complex can be further stabilized by the addition of a hydrophilic polymer attached to hydrophobic side chains. The complex can also be made specific for specific cells, by incorporating a targeting moiety such as an Fab′ fragment attached to a hydrophilic polymer.

Abstract: The present invention relates to a human or mammalian DNA replication origin consensus sequence which consists of a sequence selected from the group consisting of CCTMDAWKSGBYTSMAAWYWBCMYTTRSCAAATTCC (SEQ ID NO: 1); and AWMTWAAKRAWRWWKKDAVWWGAKRWWKWVWHRASSACMDWKAAKTWKGGWTWARRYWKGRKMWWTWKAWSDATAKWWWKDAKWKMWRKTT (SEQ ID NO: 4). A method for the control of initiation of mammalian DNA replication which comprises the steps of: a) inserting a consensus sequence coding for a sequence of the present invention together with a DNA fragment to form a vector capable of expression of the DNA fragment; b) introducing the vector of step a) into mammalian cells in vitro.

Abstract: The present invention relates, in general, to methods of stimulating phagocytosis and thereby combating infection and/or modulating immune complex disease, in particular, to methods of modulating the number and type of Fc receptors present on cells that normally possess such receptors, including monocytes and macrophages, as well as on cells that normally do not possess Fc receptors, such as fibroblasts, and to compounds and compositions suitable for use in such methods.

Abstract: A multifunctional molecular complex for the transfer of a nucleic acid composition to a target cell is provided The complex is comprised of A) said nucleic acid composition and B) a transfer moiety comprising 1) one or more cationic polyamines bound to said nucleic acid composition, 2) one or more endosome membrane disrupting components attached to at least one nitrogen of the polyamine and 3) one or more receptor specific binding components.

Abstract: Methods and compositions for producing a mammal capable of expressing an exogenously supplied gene in cells of the airway are disclosed. Liposome-nucleic acid complexes are prepared then delivered via aerosol to the lung airway. The invention provides a direct method for transforming pulmonary cells, treat disorders of the lung, and for delivering substances systematically following expression in the lung.

Abstract: The present invention provides mammalian retinoblastoma (Rb) protein-interacting zinc finger (RIZ) proteins, nucleic acid molecules encoding the RIZ and antibodies specific for the RIZ. The invention also provides screening assays for identifying an agent that effectively alters the association of a RIZ with a second molecule, which can bind to the RIZ. The invention also provides active fragments of RIZ containing the PR domain, which can regulate transcription. In addition, the invention provides methods for introducing a nucleic acid molecule-encoding a RIZ into a cell and for A-contacting a cell with an effective agent in order to modulate a function of a cell. Such methods are useful, for example, for inducing growth of a cardiac cell or a neuronal cell in a subject and for effecting normal growth control to a tumor cell or causing differentiation of tumor cells. The invention further provides methods for detecting a RIZ in a sample by detecting the RIZ or a nucleic acid molecule encoding the RIZ.

Abstract: The invention relates to new lipid compounds of formula:

Abstract: The present invention relates to a method of transfecting cells comprising applying cells directly onto nucleic acids which are immobilized in transfection complexes on a surface and which transfect the cells. Preferably, the nucleic acids are immobilized in an array. In another aspect of the present invention, the method further includes expression of the nucleic acids in the transfected cells. In yet another aspect of the present invention, the method further comprises detecting the expression of the nucleic acids in the transfected cells.

Abstract: Processes for making neutral or anionic complexes containing sequestered DNA for gene transfer, by forming a stable colloid containing an aqueous phase having suspended therein a first DNA complex with a cationic surface potential comprising a DNA sequence complexed with a cationic lipid or polymer, and modifying the surface potential of the first DNA complex to form a stable colloid comprising a second DNA complex with a neutral or net anionic surface potential.

Abstract: Methods and compositions for delivering pharmaceutical agents into cells, in particular urothelial cells of the bladder, are provided. In the methods and compositions of the invention, a solubilized cholesterol composition is used to facilitate delivery of pharmaceutical agents. Preferably, the cholesterol is solubilized by a cyclodextrin (e.g., methyl-&bgr;-cyclodextrin) and the pharmaceutical agent comprises a polynucleotide and either a cationic lipid, a cationic polymer or a dendrimer. Improved methods for transfecting polynucleotides into cells thus are also provided, using cationic lipids, cationic polymers or dendrimers and solubilized cholesterol, wherein the transfection efficiency is enhanced compared to use of cationic lipids, cationic polymers or dendrimers alone.

Abstract: The present invention relates to compositions and methods for transferring nucleic acids into cells in vitro and in vivo. The compositions comprise a transfection reagent and one or more detergents. In preferred embodiments, the compositions comprise delivery systems providing nucleic acid transfer complexes that transfect cells with high efficiency.

Abstract: The present invention relates to methods of immortalizing various primary cell cultures, including pituitary cells, neurons, beta islet cells, glial cells, corneal epithelial cells and follicular stellate cells. The primary cells are transfected with a vector containing an establishment oncogene, resulting in non-transformed immortalized cells. The primary cells and/or the subsequent immortalized cells are cultured in a defined media containing one or more environmental factor(s) that control the proliferation and/or differentiation of the cells.

Abstract: A complex that comprises (i) a nucleic acid, (ii) an integrin-binding component, for example, an integrin-binding peptide, (iii) a polycationic nucleic acid-binding component, for example, oligolysine, and (iv) a lipid component, for example, a cationic liposome, has transfection activity.

Abstract: The invention is a drug delivery system of a delivery vehicle having a low molecular weight hyaluronan ligand with an affinity for CD44 receptors. Preferably, the delivery vehicle is a liposome but other suitable delivery vehicles include microspheres, micelles, emulsions, lipid discs, polymers, viral particles and viruses. The systems of the invention may further comprise a drug, which can be any anticancer agent or other therapeutic or diagnostic agent. The invention also comprises methods of delivering a drug to a cell that expresses CD44 by contacting the cell with the drug delivery system. Further methods include treating a patient with cancer and targeting drug delivery to cells that express CD44 by attaching a glycosaminoglycan ligand.

Abstract: A composition for in vivo transfection of vertebrate male germ cells comprises a nucleic acid or transgene, and a gene delivery system, and optionally a protective internalizing agent, such as an endosomal lytic agent, a virus or a viral component, which is internalized by cells along with the transgene and which enhances gene transfer through the cytoplasm to the nucleus of the male germ cell. A pharmaceutical preparation and a transfer kit utilize the composition. A method for introducing a polynucleotide into vertebrate male germ cells comprises the administration of the composition to a vertebrate. A method for isolating or selecting transfected cells utilizes a reporter gene, and a method for administering transfected male germ cells utilizes male germ cells which have been transfected in vitro.

Abstract: A composition for in vivo transfection of vertebrate male germ cells comprises a nucleic acid or transgene, and a gene delivery system, and optionally a protective internalizing agent, such as an endosomal lytic agent, a virus or a viral component, which is internalized by cells along with the transgene and which enhances gene transfer through the cytoplasm to the nucleus of the male germ cell. A pharmaceutical preparation and a transfer kit utilize the composition. A method for introducing a polynucleotide into vertebrate male germ cells comprises the administration of the composition to a vertebrate. A method for isolating or selecting transfected cells utilizes a reporter gene, and a method for administering transfected male germ cells utilizes male germ cells which have been transfected in vitro.

Abstract: The present invention provides methods for inhibiting proliferation of astrocytes and astrocytic tumor cells. The present invention further provides methods for treating a condition associated with a defect in astrocyte proliferation in a subject, and methods for treating a condition associated with astrocytic tumor cell proliferation in a subject. Additionally, the present invention is directed to pharmaceutical compositions comprising CD81 protein or nucleic acid and a pharmaceutically-acceptable carrier. Finally, the present invention provides a method for determining whether a subject has an astrocytoma, and a method for assessing the efficacy of astrocytoma therapy in a subject.

Abstract: The present invention is directed to a Can1 mammalian sequence. Defects in this sequence result in aberrant migration and/or proliferation of primordial germ cells during embryonic development, leading to Sertoli Cell Only syndrome in males and Premature Ovarian Failure in females.

Abstract: The present invention provides a method for enhancing the level of perfusion of blood to a nonischemic skeletal muscle, e.g., by inducing angiogenesis or collateral blood formation in a nonischemic skeletal muscle at risk of being affected by ischemia or a vascular occlusion, by administration of a pharmaceutical composition comprising a DNA encoding an angiogenic peptide, such that the level of perfusion of blood to the nonischemic skeletal muscle is enhanced.

Abstract: Compositions for stabilizing polynucleic acids and increasing the ability of polynucleic acids to cross cell membranes and act in the interior of a cell. In one aspect, the invention provides a polynucleotide complex between a polynucleotide and certain polyether block copolymers. The polynucleotide complex can further include a polycationic polymer, as well as suitable targeting molecules and surfactants. The invention also provides a polynucleotide complex between a polynucleotide and a block copolymer comprising a polyether block and a polycation block.

Abstract: A gene delivery system which. is both safe and results in long-term expression throughout the brain has been developed. A lipid-entrapped, polycation-condensed DNA (LPD) system has been developed for brain gene delivery, using an adeno-associated vial. (“AAV”) vector in which the transcription unit is flanked by the 145 bp inverted terminal repeats (ITR) of the adeno-associated virus. This AAV plasmid is more effective than a non-ITR containing plasmid in vivo. The results show that the LPD-AAV plasmid complexes efficiently transduce neurons and that gene expression can persist for over 10 months in the brain. Furthermore, the intraventricular delivery method with systemic hyperosmolality results in global gene delivery. The examples show that expression of the human aspartoacyclase (“ASPA”) gene in children with this metabolic disorder can be obtained over a period of many months to a year, with functional activity.

Abstract: The present invention deals with gene therapy for treating chronic heart failure and other cardiac disease states which are accompanied by a reduced number or functioning of myocardial beta-adrenergic receptors (&bgr;-AR). &bgr;-AR receptor function is augmented in transgenic animals by delivery and expression of a beta-2-adrenergic receptor gene or a gene encoding a beta adrenergic receptor kinase inhibitor, resulting in increased in vivo left ventricular function. The present invention includes recombinant plasmid vectors, alternative beta-adrenergic receptor gene delivery strategies, and transgenic mice carrying a &bgr;-AR transgene, a &bgr;-ARK transgene, or a &bgr;-ARK inhibitor transgene.

Abstract: A complex is described for delivery to a cell comprising inserting a nucleic acid into a reverse micelle. The reverse micelle has the property to compact the nucleic acid for easier delivery. Other molecules are used to interact with the nucleic acid—micelle complex to further enhance delivery such as a surfactant having a disulfide bond.

Abstract: The present invention concerns the discovery that proteins encoded by a family of vertebrate genes, termed here signalin-related genes, which are involved in signal transduction induced by members of the TGF&bgr; superfamily. The present invention makes available compositions and methods that can be utilized, for example to generate and/or maintain an array of different vertebrate tissue both in vitro and in vivo.

Abstract: An antisense oligonucleotide characterized in that it hybridizes specifically with chromosomal DNA and/or RNA encoding CXCR4 protein to thereby inhibit the expression of the CXCR4 protein, and a HIV infection inhibitor comprising the antisense oligonucleotide, are disclosed.

Abstract: A method of liposome-based therapy for a mammalian subject is disclosed. The method uses liposomes with outer surfaces that contain an affinity moiety effective to bind specifically to a biological surface at which the therapy is aimed, and a hydrophilic polymer coating effective to shield the affinity moiety from interaction with the target surface. The hydrophilic polymer coating is made up of polymer chains covalently linked to surface lipid components in the liposomes through releasable linkages. After a desired liposome biodistribution is achieved, a releasing agent is administered to cause cleaving of a substantial portion of the releasable linkages in the liposomes, to expose the affinity agent to the target surface.

Abstract: The invention is directed to a method for treating both the early and late phases of allergic asthma by introducing naked polynucleotides which operatively encode for the asthma-initiating antigen into the host. The antigen-encoding polynucleotides are administered to host tissues which contain a high concentration of antigen presenting cells (e.g., skin and mucosa) relative to other host tissues. Expression of the asthma-initiating antigen encoding polynucleotides of the invention inside of antigen presenting cells (without substantial secretion therefrom) induces antigen tolerance while suppressing IgE antibody formation in the early phase of the disease, and also suppresses cytokine-mediated eosinophil accumulation in the late phase of the disease. Devices and compositions for use in the methods of the invention are also described.

Abstract: The invention relates to a novel CFTR gene regulatory element capable of increasing the activity of the CFTR gene promoter, and to nucleic acid constructs comprising the element together with the CFTR gene coding region. The element and constructs containing it are useful in gene therapy for treating cystic fibrosis.

Abstract: The present invention relates to a method for introducing one or more antisense oligonucleotides into one or more eucaryotic cells using one or more lipid formulations comprising one or more cationic lipids of Formula I and optionally at least one neutral lipid. In particular, the present invention relates to a method for introducing one or more antisense oligonucleotides into one or more eucaryotic cells using a lipid formulation comprising dimethyldioctadecylammonium bromide (DDAB) and at least one neutral lipid, especially dioleylphosphatidylethanolamine (DOPE). The invention also relates to kits for carrying out the invention, compositions for carrying out the invention, and compositions formed while carrying out the invention. Further, the present invention relates to a method for inhibiting or preventing cell growth or proliferation, and a method for inhibiting or preventing expression of one or more proteins.

Abstract: A protein, designated ERCoA3 is provided. The ERCoA3 protein interacts with the estrogen receptor and the progesterone receptor and causes activation of these receptors is provided. Also provided are polynucleotides which encode ERCoA3 or block translation of the mRNA which encodes ERCoA3. Antibiodies that bind to one or more epitopes in the human ERCoA3 protein are provided. The present invention also relates to methods of inhibiting or reducing tamoxifen or estrogen induced proliferation of cancer cells, particularly breast cancer cells, endometrial cancer cells and uterine cancer cells. The method comprises reducing the activity or levels of ERCoA3 in such.

Abstract: Novel cationic polymers and cationic lipids, and methods of making and using the same, are provided. The cationic polymers and cationic lipids are useful for the delivery of nucleic acid polymers and oligomers to cells in vitro and in vivo.

Abstract: The present invention provides an isolated nucleic acid covalently linked to a photolabile caging group which reversibly prevents expression of the nucleic acid. The present invention further provides a method of selectively expressing a nucleic acid in a cell, comprising: a) covalently linking the nucleic acid to a photolabile caging group which reversibly prevents expression of the nucleic acid; b) introducing the nucleic acid of step (a) into the cell; and c) exposing the cell of step (b) to light, whereby exposure to the light unlinks the nucleic acid and the caging group and the nucleic acid is selectively expressed in the cell.

Abstract: The present invention relates to methods and compositions for sensitizing a cell to a prodrug compound.

Abstract: A gene delivery system which is both safe and results in long-term expression throughout the brain has been developed. A lipid-entrapped, polycation-condensed DNA (LPD) system has been developed for brain gene delivery, using an adeno-associated viral (“AAV”) vector in which the transcription unit is flanked by the 145 bp inverted terminal repeats (ITR) of the adeno-associated virus. This AAV plasmid is more effective than a non-ITR containing plasmid in vivo . The results show that the LPD-AAV plasmid complexes efficiently transduce neurons and that gene expression can persist for over 10 months in the brain. Furthermore, the intraventricular delivery method with systemic hyperosmolality results in global gene delivery. The examples show that expression of the human aspartoacyclase (“ASPA”) gene in children with this metabolic disorder can be obtained over a period of many months to a year, with functional activity.

Abstract: A method for immunization using genetic material is disclosed. Compositions for genetic immunization comprising cationic lipids and polynucleotides are also disclosed. Methods for using genetic immunization to produce polyclonal and monoclonal antibodies are also disclosed. A method for epitope mapping is also disclosed.

Abstract: The present invention provides novel and surprisingly effective methods for delivering nucleic acids to cells. These methods are based upon the discovery that the presence of endosomal membrane destabilizers (e.g., calcium) leads to a dramatic increase in the transfection efficiency of plasmids formulated as SPLP, or “stabilized plasmid-lipid particles.