Abstract: The present invention provides biodegradable polymer nanospheres capable of transporting and releasing therapeutic agents, specifically nucleic acids. In preferred embodiments, a sub-150 nm nanosphere is formed containing nucleic acids. Thereafter, the agent is released from the nanosphere. In one embodiment a biodegradable polymer nanosphere surface has attached to it a targeting moiety. In another embodiment, the biodegradable polymer nanosphere surface has attached to it a masking moiety. In yet another embodiment both targeting and masking moieties are attached to the nanosphere surface.

Abstract: An improved particle bombardment device for transporting biological substances such as DNA into living cells. The device has a flexible barrel (40) that facilitates endoscopic particle bombardment of in vivo cells without a significant concomitant blast effect and without a need for a vacuum. The device also involves a unique tapered particle-carrying macroprojectile that can travel through convolutions of such a flexible barrel (40) with minimal friction.

Abstract: A method for transfecting and separating cells is disclosed. The method comprises preparing magnetic particles coated with genetic material and a cell-specific ligand, and using the particles to transfect target cells. The target cells may then be separated from the non-target cells by using a magnetic field.

Abstract: The present invention is directed to the method and apparatus for the cytoplasmic loading of macromolecules into living cells by an impact-mediated procedure that impacts the cells with a predetermined number of solid particles in a blast of propellant gas. More specifically, the present invention is directed to an impact-mediated procedure that is altered by gravitational conditions and is preferably carried out under hypergravity conditions. Further, the present invention is directed to an IML method and apparatus for consistently and reproducibly loading macromolecules into the cytoplasm of living cells via membrane wounding at significantly higher efficiencies than can be accomplished using existing methodologies. The IML procedure directs a blast of propellant gas through a rupturable membrane on which solid particles are supported in order to achieve insertion of a predetermined number of particles into the propellant blast.

Abstract: The invention relates to an ionic conjugate, which is stable in a biological medium, and which is comprised of a particle vector with at least one cationic, nonliquid, hydrophilic nucleus and of polyanionic oligonucleotides. The invention further concerns the pharmaceutical compositions containing these conjugates and the use of a particle vector to carry the oligonucleotides to the cells.

Abstract: An approach to genetic vaccine methodology is described. A genetic construction encoding antigenic determinants of a filovirus is transfected into cells of the vaccinated individuals using a particle acceleration protocol so as to express the viral antigens in healthy cells to produce an immune response to those antigens.

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 coding information for three putative chicken anemia virus proteins (VP1, VP2, VP3) was inserted into a baculovirus vector and expressed in insect cells. The immunogenic properties of the chicken anemia virus (CAV) proteins produced separately or together in insect-cell cultures were analyzed by inoculating them into chickens. Only lysates of insect cells which have synthesized equivalent amounts of all three recombinant CAV proteins or cells which synthesized mainly VP1 plus VP2 induced neutralizing antibodies directed against CAV in inoculated chickens. Progeny of those chickens were protected against clinical disease after CAV challenge. Inoculation of a mixture of lysates of cells that were separately infected with VP1-, VP2- and VP3-recombinant baculovirus did not induce significant levels of neutralizing antibody directed against CAV and their progeny were not protected against CAV challenge.

Abstract: A method of introducing a biological substance into living target cells, the method comprising dispersing a liquid containing the biological substance into microdroplets and propelling the microdroplets toward to the target cells.

Abstract: The present invention relates to a method of transforming cells in which an appropriate quantity of nucleic acid fragments is introduced into the cells. The nucleic acid fragments are introduced in the form of a nucleic acid composition comprising a nitrogen-containing silicone, useful for compacting the nucleic acid fragments; the compositions comprise aggregates of nucleic acid fragments and silicones according to the invention.

Abstract: The present invention relates to transfected primary and secondary somatic cells of vertebrate origin, particularly mammalian origin, transfected with exogenous genetic material (DNA) which encodes a desired (e.g., a therapeutic) product or is itself a desired (e.g., therapeutic) product, methods by which primary and secondary cells are transfected to include exogenous genetic material, methods of producing clonal cell strains or heterogenous cell strains, methods of gene therapy in which the transfected primary or secondary cells are used, and methods of producing antibodies using the transfected primary or secondary cells.

Abstract: The present invention relates to methods and compositions for delivery of a compound, most preferably a polynucleotide, into the cytoplasm of a cell by means of a microparticle, fabricated from a pH-sensitive hydrogel in collapsed phase, and having a size and physical characteristics compatible with uptake via a clathrin-coated pit on the cell surface. The trigger pH at which the hydrogel expands is lower than the physiological pH of the extracellular environment, allowing the microparticle, which comprises the compound to be delivered, to maintain its compact size prior to cellular uptake, thereby providing the additional benefit of protecting the comprised compound from degradation. This protective feature is particularly beneficial in embodiments in which a polynucleotide or a peptide is being delivered. Following uptake via a clathrin-coated pit, the microparticle then enters the intracellular endocytic pathway, where it is subjected to a progressive decrease in pH.

Abstract: In an approach to genetic therapy for treating tumors, a genetic construction encoding the p35 and p40 subunits of the cytokine IL-12 is delivered into cells of individuals in need of therapy so as to express IL-12 in cells and to cause regression of established tumors.

Abstract: The present invention describes a reliably protective avian vaccine that contains a DNA transcription unit comprising a nucleic acid that encodes a viral protein. This effective and reliable avian vaccine significantly improves upon that which heretofore has been disclosed. The present invention demonstrates that the reliable efficacy found for DNA influenza vaccines, formerly observed in experimental inbred mice, can now be obtained for outbred avian species as well. Methods of use and specific DNA constructs are disclosed.

Abstract: This invention relates to an isolated and purified DNA encoding an amino acid sequence which comprises SEQ. ID. NO:3.

Abstract: In a tubular pressure chamber a drop of a DNA solution with gold particles suspended therein is atomized at the opening of a cannula by a pressure impact. The fog droplets containing the gold particles and the DNA entrained by them, are pressured by the pressure impact through a restriction at the end of the pressure chamber and thereby accelerated and focused. They subsequently traverse in free flight an evacuated specimen chamber and impact in a narrowly limited target area with a predetermined pulse, cells fixed on a holder, thereby penetrating them.