Abstract: The present invention provides dynamic charge state cationic polymers that are useful for delivery of anionic molecules. The dynamic charge state cationic polymers are designed to have cationic charge densities that decrease by removal of removable functional groups from the polymers. The present invention also provides interpolyelectrolyte complexes containing the polymers complexed to a polyanion. Methods for using the interpolyelectrolyte complexes to deliver anionic compounds are also provided.

Abstract: Electroporation is performed in a controlled manner in either individual or multiple biological cells or biological tissue by monitoring the electrical impedance, defined herein as the ratio of current to voltage in the electroporation cell. The impedance detects the onset of electroporation in the biological cell(s), and this information is used to control the intensity and duration of the voltage to assure that electroporation has occurred without destroying the cell(s). This is applicable to electroporation in general. In addition, a particular method and apparatus are disclosed in which electroporation and/or mass transfer across a cell membrane are accomplished by securing a cell across an opening in a barrier between two chambers such that the cell closes the opening. The barrier is either electrically insulating, impermeable to the solute, or both, depending on whether pore formation, diffusive transport of the solute across the membrane, or both are sought.

Abstract: Reagents useful in nucleic acid immunization techniques are described. More particularly, adjuvanted genetic vaccine compositions are described, as are methods of using those compositions for inducing an enhanced immune response against a selected antigen.

Abstract: Methods and composition of induction of pluripotent stem cells are disclosed. For example, in certain aspects methods for generating induced pluripotent stem cells using reporter genes are described. Furthermore, the invention provides novel reprogramming vectors employing reporter genes.

Abstract: The present invention relates to a novel crosslinked polyethylenimine (PEI) nanoparticle based nucleic acid transfection agent wherein the crosslinker is having carbon chain in the range of C2 to C8, ranging between 3.27-19.8%, having the size of nanoparticle ranging between 20-600 nm and zeta potential ranging from +5 to 50 mV.

Abstract: There is provided a method by which multiple types of substances desired to be transferred into cells can be continuously transferred into multiple types of cells by a convenient procedure, a cell in which the substance desired to be transferred into cells has been taken up by this method, and an apparatus for transferring a substance into cells by this method. The foregoing objects can be achieved by electrospraying cells with a liquid free from the substance to be transferred into cells while the cells are kept in contact with the substance to be transferred into cells, or first electrospraying cells with a liquid free from the substance to be transferred into cells and then bringing the cells into contact with the substance to be transferred into cells.

Abstract: Disclosed are methods and compositions to genetically modify substantially intact cells having cosmetic function to enhance the cosmetic appearance in mammals so as to enhance and/or maintain a biochemical and/or physiological process that has a positive effect on cosmetic appearance. The methods and compositions may provide cosmetic benefits such as reduced skin sagging, increased skin thickness, reduced wrinkles, increased skin thickness and collagen content, increased skin tone and elasticity, increased skin hydration, and improved skin texture and color.

Abstract: The present invention provides methods and compositions useful in localized transfer of genetic material or proteins. Moreover, the present invention provides methods and compositions for improving and/or controlling wound healing by applying a wound care device comprising HoxD3 and/or HoxA3 and/or HoxB3. In addition, the present invention provides methods and compositions for improved wound healing in subjects having impaired healing capabilities, such as diabetic and aged subjects.

Abstract: The invention provides a method of activating hepatitis C virus (HCV)-specific T cells, including CD4+ and CD8+ T cells. HCV-specific T cells are activated using fusion proteins comprising HCV NS3, NS4, NS5a, and NS5b polypeptides, polynucleotides encoding such fusion proteins, or polypeptide or polynucleotide compositions containing the individual components of these fusions. The method can be used in model systems to develop HCV-specific immunogenic compositions, as well as to immunize a mammal against HCV.

Abstract: The invention provides transgenomic mitochondria, transmitochondrial cells and organisms, and the materials and methods for making such mitochondria, cells, and organisms.

Abstract: The invention relates to compositions and methods for reverse gene therapy, wherein a gene therapy vector encoding a gene product (e.g. a protein) which is usually only expressed in cells of an abnormal tissue is delivered to a cell of an animal afflicted with a disease or disorder to alleviate the disease or disorder. In one embodiment, a plasmid vector encoding HERG (A561V) protein is delivered to a cell of an animal afflicted with re-entrant atrial flutter-mediated cardiac arrhythmia.

Abstract: A method of using vaults as carrier molecules to deliver one or more than one substance to an organism, or to a specific tissue or to specific cells, or to an environmental medium. A vault-like particle. A method of preventing damage by one or more than one substance to an organism, to a specific tissue, to specific cells, or to an environmental medium, by sequestering the one or more than one substance within a vault-like particle. A method of delivering one or more than one substance or a sensor to an organism, to a specific tissue, to specific cells, or to an environmental medium. According to another embodiment of the present invention, there is provided a method of making vault-like particles, and making vault-like particles comprising one or more than one substance, or one or more than one sensor.

Abstract: Superior molecular vaccines comprise nucleic acids, including naked DNA and replicon RNA, that encode a fusion polypeptide that includes an antigenic peptide or polypeptide against which an immune response is desired. Fused to the antigenic peptide is an intercellular spreading protein, in particular a herpes virus protein VP22 or a homologue or functional derivative thereof. Preferred spreading proteins are VP22 from HSV-1 and Marek's disease virus. The nucleic acid can encode any antigenic epitope of interest, preferably an epitope that is processed and presented by MHC class I proteins. Antigens of pathogenic organisms and cells such as tumor cells are preferred. Vaccines comprising HPV-16 E7 oncoprotein are exemplified. Also disclosed are methods of using the vaccines to induce heightened T cell mediated immunity, in particular by cytotoxic T lymphocytes, leading to protection from or treatment of a tumor.

Abstract: A method of transferring a gene to vertebrate cells is disclosed. The method comprises the steps of: (a) providing microprojectiles, the microprojectiles carrying polynucleic acid sequences, the sequences comprising, in the 5? to 3? direction, a regulatory sequence operable in the tissue cells and a gene positioned downstream of the regulatory sequence and under the transcriptional control thereof; and (b) accelerating the microprojectiles at the cells, with the microprojectiles contacting the cells at a speed sufficient to penetrate the cells and deposit the polynucleic acid sequences therein. Preferably, the target cells reside in situ in the animal subject when they are transformed. Preferred target cells are dermis or hypodermis cells, and preferred genes for insertion into the target cells are genes which code for proteins or peptides which produce a physiological response in the animal subject.

Abstract: Different-sequence peptoids, including lipid- and sterol-conjugated peptoids, are found to be effective in transfection of cells with oligonucleotides. Combinatorial libraries of such peptoids can be screened efficiently in a high-throughput format, and selected peptoids are identified post-screening.

Abstract: A method of highly efficiently transferring various selected molecules into various cells and a method of fusing cells. Cells and/or selected molecules such as polynucleotide are treated with cold gas plasma to thereby transfer the selected molecules located around cells into the cells, or cells are fused by treating the cells with cold gas plasma. Moreover, an apparatus for transferring selected molecules or fusing cells having a cold gas plasma generation unit for transferring selected molecules into cells is provided.

Abstract: The invention relates to the use of DNA-loaded nanoparticles for a transfection of DNA into cells in vitro or in vivo.

Abstract: The present invention includes compositions and methods for transforming cells into glucose-responsive, insulin-production cells using a construct that expresses betacellulin and PDX1, e.g., transforming pancreatic acinar cells using one or more expression vectors that expressed betacellulin and PDX1 using ultrasound targeted microbubble destruction (UTMD).

Abstract: A method of transferring a gene to vertebrate cells is disclosed. The method comprises the steps of: (a) providing microprojectiles, the microprojectiles carrying polynucleic acid sequences, the sequences comprising, in the 5? to 3? direction, a regulatory sequence operable in the tissue cells and a gene positioned downstream of the regulatory sequence and under the transcriptional control thereof; and (b) accelerating the microprojectiles at the cells, with the microprojectiles contacting the cells at a speed sufficient to penetrate the cells and deposit the polynucleic acid sequences therein. Preferably, the target cells reside in situ in the animal subject when they are transformed. Preferred target cells are dermis or hypodermis cells, and preferred genes for insertion into the target cells are genes which code for proteins or peptides which produce a physiological response in the animal subject.

Abstract: Disclosed are methods and compositions to genetically modify substantially intact cells having cosmetic function to enhance the cosmetic appearance in mammals so as to enhance and/or maintain a biochemical and/or physiological process that has a positive effect on cosmetic appearance. The methods and compositions may provide cosmetic benefits such as reduced skin sagging, increased skin thickness, reduced wrinkles, increased skin thickness and collagen content, increased skin tone and elasticity, increased skin hydration, and improved skin texture and color.

Abstract: The present invention relates to a process for inducing a protective immune response in a mammal against a virus, in which a construct including a promoter and a DNA sequence encoding a protein or peptide producing an immune response against the virus is introduced into muscle or skin of the mammal via a microprojectile. Sufficient amounts of the construct are administered so as to produce a protective immune response in the mammal.

Abstract: This invention is predicated on the present applicants' discovery that nanostructures comprising discrete regions of different composition can be used to deliver to a biological cell a desired combination of molecules in close proximity. Different molecules can be selectively bonded to discrete regions of different composition in sufficiently close physical relationship to enhance delivery or effectiveness within the cell. The preferred nanostructures are multicomponent nanorods. Important applications include delivery of missing DNA sequences for gene therapy and delivery of antigens or DNA encoding antigens for vaccination.

Abstract: Porous and/or polycrystalline silicon are used in the delivery of substances into cells. The porous and/or polycrystalline silicon can be formed into micropiercers, microneedles and biolistic bullets for penetration of the cell. The control of the pore size and porosity of the porous and/or polycrystalline silicon allows tuning of the bioactivity of the porous silicon. The porous and/or polycrystalline silicon is also resorbable and is therefore resorbed from the cells without leaving any particles or being seen as a foreign body. Methods of manufacturing the porous silicon micropiercers, microneedles, microelectrodes, biolistic bullets, and precipitation of calcium phosphate on a bioactive substrate, and their advantages over known methods of delivering materials into cells.

Abstract: An electrospraying apparatus and/or method is used to coat particles. For example, a flow including at least one liquid suspension may be provided through at least one opening at a spray dispenser end. The flow includes at least particles and a coating material. A spray of microdroplets suspending at least the particles is established forward of the spray dispenser end by creating a nonuniform electrical field between the spray dispenser end and an electrode electrically isolated therefrom. The particles are coated with at least a portion of the coating material as the microdroplet evaporates. For example, the suspension may include biological material particles.

Abstract: In order to ensure consistently good maceration of biological material in an electroporation reactor, it is proposed to monitor the conductivity of the mixture therein and to detect any arcing which occurs therein. The results of such monitoring are used to modify the operating voltage of the electroporation reactor and/or the composition of the mixture located in the reactor channel.

Abstract: The present invention relates to various methods of genetic immunization for the purpose of providing antigen-specific immunity in a mammalian host, including a human host. The invention is based on the ability to direct particulate polynucleotides which express an antigenic protein or protein fragment to the cytoplasm of host target cells, such as antigen presenting cells. A directed delivery of such particulate polynucleotides to the cytoplasm of antigen presenting cells will stimulate antigen-specific CTL production, thus promoting destruction of affected cells such as neoplastic cells and virally infected cells.

Abstract: The present invention provides methods and compositions useful in localized transfer of genetic material or proteins. Moreover, the present invention provides methods and compositions for improving and/or controlling wound healing by applying a wound care device comprising HoxD3 and/or HoxA3 and/or HoxB3. In addition, the present invention provides methods and compositions for improved wound healing in subjects having impaired healing capabilities, such as diabetic subjects.

Abstract: A method for introducing a foreign matter into a cell, includes the steps of placing a small particle carrying a foreign matter at a part of a cell surface of a living cell, boring a hole in a cell wall and/or a cell membrane by irradiating and treating said part of the cell surface with a laser beam, and introducing the foreign matter into the living cell.

Abstract: The present invention provides methods and compositions useful in localized transfer of genetic material or proteins. Moreover, the present invention provides methods and compositions for improving and/or controlling wound healing by applying a wound care device comprising HoxD3 and/or HoxA3. In addition, the present invention provides methods and compositions for improved wound healing in subjects having impaired healing capabilities, such as diabetic subjects.

Abstract: An ordered array of magnetized nanorods includes a plurality of metallic nanorods generally cylindrical in shape and including a nickel portion coated with a positively charged polyelectrolyte and a gold portion coated with an alkanethiolate; and a layer of a hardened polymer wherein each individual nanorod of the plurality is held by having said gold portion embedded therein so that the nickel portion extends approximately perpendicularly away from the layer of hardened polymer, and wherein said plurality of metallic nanorods is ordered in the array by having substantially all individual nanorods of the plurality of nanorods oriented generally parallel to each other.

Abstract: The present invention relates to the use of fluorogenic or chromogenic dyes as reporter molecules for detecting cell entry by a specific molecule.

Abstract: The invention relates to a particulate product comprising at least one microprojectile; characterized in that the or at least one of the microprojectiles comprises silicon. The invention also relates to devices and components used in the microprojectile implantation of the particulate product to a target of cells or target tissue.

Abstract: A novel garden bean cultivar, designated ‘210104’, is disclosed. The invention relates to the seeds of garden bean cultivar ‘210104’, to the plants of garden bean line ‘210104’ and to methods for producing a bean plant by crossing the cultivar ‘210104’ with itself or another bean line. The invention further relates to methods for producing a bean plant containing in its genetic material one or more transgenes and to the transgenic plants produced by that method and to methods for producing other garden bean lines derived from the cultivar ‘210104’.

Abstract: The inventive vector specifically directs entry into a cell of monocytic origin. The vector is composed of a nucleic acid component, a lysosome evading component and a particle that can be phagocytized. The vector itself, or cells pretreated with the vector, are useful in all gene medicine applications. Because it is specific for monocytic cells, the inventive vector is particularly suited to vaccine applications. Due to the ability of monocytic cells to target tumors, the inventive vector also is suitable for use in anti-tumor applications, including conventional gene therapy.

Abstract: A novel potato cultivar of the genus and species Solanum tuberosum, designated FL1900, is disclosed. The invention relates to the tubers of potato variety FL1900, to the plants of potato variety FL1900, to the seeds of potato variety and to methods for producing hybrid potato variety. The invention further relates to potato variety tubers, seeds and plants produced by crossing the potato variety FL1900 with another potato plant, and to Single Gene Converted plants.

Abstract: A method is provided for the labeling of individual cells. Labeling is accomplished by coating a particle with at least one dye or nucleic acid sequence encoding a marker protein. The particle is then propelled toward the cell resulting in the particle contacting the cell for a time sufficient for the dye or nucleic acid to leave the particle and enter the cell. The present method allows for the differential labeling of individual cells within dense populations of cells.

Abstract: The invention provides primary and secondary cells that are transfected with a nucleic acid molecule that encodes erythropoietin, clonal or heterogenous strains of such cells, and methods of producing these cell strains.

Abstract: The present invention discloses a general method for the efficient delivery of suppressor tRNA and mRNA or DNA encoding a protein of interest to mammalian cells. The method of electroporation described herein is useful for expanding nonsense suppression methodology to mammalian cell expression systems for the incorporation of unnatural amino acids.

Abstract: Recombinant antibody-based molecules that trigger both T-cell and B-cell immune responses are disclosed. The recombinant molecules are comprised by at least one targeting unit and at least one antigenic unit connected through a dimerization motif. Also disclosed are nucleic acid molecules encoding the recombinant antibody-based molecule and methods of treating multiple myeloma or lymphoma in a patient using the recombinant antibody-based molecules or the nucleic acid molecules.

Abstract: The controllable electroporation system and method described herein allows control over the size, the number, the location, and the distribution of aqueous pores, thus increasing flexibility of use. The herein described system and method for controllable electroporation generally employs at least two actuating sub-systems and sub-processes. One sub-system and sub-process employs a relatively broad effect in order to weaken the membrane, a broad effect sub-system. Another sub-system and sub-process employs a relatively narrow effect in order to localize the position of the pore in the membrane, a narrow effect sub-system.

Abstract: The present invention relates to a method of inserting RNA into cells. In this method, cells are transfected with RNA using electroporation in order to achieve high transfection efficiency. The method is useful, inter alia, in providing cells to be used in cell-based therapies, e.g. in preparing cells useful as anti-cancer vaccines. Preferably, the RNA has a 5′ cap and a 3′ poly (A) tail.

Abstract: A method is provided for introducing nucleic acid into a cell, by contacting the cell with a nucleic acid and applying a low electrical field impulse for a long pulse length. A method is provided for introducing a polypeptide into a cell, by contacting the cell with the polypeptide and applying a low electrical field impulse for a long pulse length.

Abstract: Therapeutic compositions are delivered to a target site using a catheter which includes at least one ultrasound transducer coupled to an energy source. The therapeutic compositions include genetic material and the target site may be a DNA with affinity for the genetic material.

Abstract: Methods for loading an antigen-presenting cell with one ore more antigens are disclosed. Methods for the treatment and prevention of a disease in a subject using an antigen-presenting cell that has been electroporated with a composition of one or more antigens. Composition of one or more antigens comprises one or more antigens of a hyperproliferative cell, a microorganism or a microorganism-infected cell are also disclosed. In addition, compositions of antigen-presenting cells that have been loaded with one or more antigens of a hyperproliferative cell, a microorganism-infected cell or a microorganism using electroporation are disclosed.

Abstract: Systems and methods are described for parallel macromolecular delivery and biochemical/electrochemical interface to whole cells employing carbon nanostructures including nanofibers and nanotubes. A method includes providing a first material on at least a first portion of a first surface of a first tip of a first elongated carbon nanostructure; providing a second material on at least a second portion of a second surface of a second tip of a second elongated carbon nanostructure, the second elongated carbon nanostructure coupled to, and substantially parallel to, the first elongated carbon nanostructure; and penetrating a boundary of a biological sample with at least one member selected from the group consisting of the first tip and the second tip.

Abstract: The present invention relates to a gene gun and the application of the gene gun for gene transformation. A low pressure gas is used in the gene gun to directly accelerate the biological material containing solution, so that the biological materials penetrate through the cell membrane/wall or the skin of an animal, without using metal particle carriers, for gene transformation.

Abstract: The present invention provides a method for gene transfer through the vascular system or a ureter. In a preferable embodiment of this invention, an aqueous solution containing an expression vector comprising a desired gene is injected through the vascular system or a ureter of a desired organ to introduce the gene into the injected area. Specifically, the method of the present invention is effective for ex vivo gene therapy wherein the hepatocyte growth factor (HGF) gene is introduced by electroporation into a kidney to be transplanted. According to the present invention, the HGF functions continuously on the kidney to be transplanted, and the HGF within the treated area does not affect other organs of the body. Thus, the method of the present invention is a safe and effective treatment for maintaining the survival of transplanted kidneys.

Abstract: A method of transferring a gene to vertebrate cells is disclosed. The method comprises the steps of: (a) providing microprojectiles, the microprojectiles carrying polynucleic acid sequences, the sequences comprising, in the 5′ to 3′ direction, a regulatory sequence operable in the tissue cells and a gene positioned downstream of the regulatory sequence and under the transcriptional control thereof; and (b) accelerating the microprojectiles at the cells, with the microprojectiles contacting the cells at a speed sufficient to penetrate the cells and deposit the polynucleic acid sequences therein. Preferably, the target cells reside in situ in the animal subject when they are transformed. Preferred target cells are dermis or hypodermis cells, and preferred genes for insertion into the target cells are genes which code for proteins or peptides which produce a physiological response in the animal subject.

Abstract: The present invention relates to the use of porous silicon in the delivery of substances into cells. The porous silicon can be formed into micropiercers, microneedles and biolistic bullets for peenetration of the cell. The control of the pore size and porosity of the porous silicon allows tuning of the bioactivity of the porous silicon. The porous silicon is also resorbable and is therefore resorbed from the cells without leaving any particles or being seen as a foreign body. The present invention also relates to the methods of manufacturing the porous silicon micropiercers, microneedles, microelectrodes, biolistic bullets, and precipitation of calcium phosphate on a bioactive substrate, and their advantages over known methods of delivering materials into cells.

Abstract: Methods of killing neoplastic cells are provided. The invention relates to the use of folylpolyglutamyl synthetase (FPGS) gene transfer to enhance the sensitivity of several types of tumor cells to polyglutamylatable antifolate drugs, such as methotrexate (MTX) and edatrexate (EDX).