Abstract: Targeted non-surgical administration of a nucleic acid formulation to the suprachoroidal space (SCS) of the eye of a human subject permits effective treatment of ocular disorders, including posterior ocular or choroidal maladies. In one embodiment, the method comprises inserting a hollow microneedle into the eye at an insertion site and infusing a nucleic acid formulation through the inserted microneedle and into the suprachoroidal space of the eye. The infused nucleic acid formulation flows within the suprachoroidal space away from the insertion site. In one embodiment, the fluid nucleic acid formulation comprises nucleic acid nanoparticles consisting of one molecule of nucleic acid.

Abstract: Efficient and prolonged hCFTR expression is one of the major obstacles for cystic fibrosis lung therapy. hCFTR mRNA expression levels depend on eukaryotic expression cassette components, prokaryotic backbone elements, and the gene transfer method may also influence transcriptional silencing mechanisms. A codon-optimized and CpG-reduced human CFTR gene (CO-CFTR) was made. Various vector modifications were tested to facilitate extended duration of CO-CFTR expression. Insertion of an extended 3?BGH transcribed sequence (712 bp) in an inverted orientation produced prolonged expression of CO-CFTR expression at biologically relevant levels. Further studies revealed that prolonged CO-CFTR expression is dependant on the orientation of the extended BGH 3? BGH transcribed sequence and its transcription, is not specific to the UbC promoter, and is less dependent on other vector backbone elements.

Abstract: Counterions of polycations used to compact nucleic acids profoundly affect shape and stability of particles formed. Shape is associated with differential serum nuclease resistance and colloidal stability. A surrogate for determining such properties that is easy to measure is the turbidity parameter. Shape also affects the suitability and efficacy of compacted nucleic acid complexes for transfecting cells by various routes into a mammalian body. Moreover, counterions such as acetate can protect compacted nucleic acid complexes from adverse effects of lyophilization.

Abstract: A synthetic hCFTR DNA sequence has been developed that produces remarkably high levels of hCFTR mRNA and protein in dosed murine lungs and human cells in culture compared to the natural hCFTR cDNA. This synthetic DNA addresses problems inherent in some natural cDNAs, such as premature transcriptional truncation sites introduced during cDNA synthesis. Introns are initially present in mRNA until the mRNA is processed. cDNA made from processed mRNA is devoid of introns. Thus DNA sequences (exon junctions) are present in a cDNA molecule which are not present in cells in nature. These exon junctions may affect transcription. Methods for improving expression of CFTR are based on sequence changes in cDNA molecules. The improvement methods may be applied to other cDNA molecules which are refractory to in vivo expression efforts. Compositions embodying the sequence changes increase the production of both transgenic mRNA and protein from cDNA molecules.

Abstract: Efficient and prolonged hCFTR expression is one of the major obstacles for cystic fibrosis lung therapy. hCFTR mRNA expression levels depend on eukaryotic expression cassette components, prokaryotic backbone elements, and the gene transfer method may also influence transcriptional silencing mechanisms. A codon-optimized and CpG-reduced human CFTR gene (CO-CFTR) was made. Various vector modifications were tested to facilitate extended duration of CO-CFTR expression. Insertion of an extended 3?BGH transcribed sequence (712 bp) in an inverted orientation produced prolonged expression of CO-CFTR expression at biologically relevant levels. Further studies revealed that prolonged CO-CFTR expression is dependant on the orientation of the extended BGH 3? BGH transcribed sequence and its transcription, is not specific to the UbC promoter, and is less dependent on other vector backbone elements.

Abstract: The present invention is a method of using compacted nucleic acid (such as DNA) nanoparticles for non-viral gene transfer to various tissues of the human eye or eyes of other mammals. These nanoparticles comprise, in one embodiment, a neutrally-charged complex containing a single molecule of plasmid DNA compacted with polyethylene glycol (PEG)-substituted poly lysine peptides.

Abstract: Counterions of polycations used to compact nucleic acids profoundly affect shape and stability of particles formed. Shape is associated with differential serum nuclease resistance and colloidal stability. A surrogate for determining such properties that is easy to measure is the turbidity parameter. Shape also affects the suitability and efficacy of compacted nucleic acid complexes for transfecting cells by various routes into a mammalian body.

Abstract: Counterions of polycations used to compact nucleic acids profoundly affect shape and stability of particles formed. Shape is associated with differential serum nuclease resistance and colloidal stability. A surrogate for determining such properties that is easy to measure is the turbidity parameter. Shape also affects the suitability and efficacy of compacted nucleic acid complexes for transfecting cells by various routes into a mammalian body.

Abstract: Counterions of polycations used to compact nucleic acids profoundly affect shape and stability of particles formed. Shape is associated with differential serum nuclease resistance and colloidal stability. A surrogate for determining such properties that is easy to measure is the turbidity parameter. Shape also affects the suitability and efficacy of compacted nucleic acid complexes for transfecting cells by various routes into a mammalian body.

Abstract: Counterions of polycations used to compact nucleic acids profoundly affect shape and stability of particles formed. Shape is associated with differential serum nuclease resistance and colloidal stability. A surrogate for determining such properties that is easy to measure is the turbidity parameter. Shape also affects the suitability and efficacy of compacted nucleic acid complexes for transfecting cells by various routes into a mammalian body.

Abstract: Counterions of polycations used to compact nucleic acids profoundly affect shape and stability of particles formed. Shape is associated with differential serum nuclease resistance and colloidal stability. A surrogate for determining such properties that is easy to measure is the turbidity parameter. Shape also affects the suitability and efficacy of compacted nucleic acid complexes for transfecting cells by various routes into a mammalian body.

Abstract: An automated nucleic acid compaction device for analyzing and compacting a nucleic acid complex into unimolecular nucleic acid particles. Broadly, the device includes a container support and agitation system; a measuring and testing system; and a dispensing system; all controlled by a control system. The control system controls the support and agitation system and the dispensing system based either on a predetermined formulation or by analysis of feedback data provided by the measuring and testing system. In a preferred embodiment, the device is a real-time measuring and mixing instrument operating in a closed loop system. The preferred embodiment also comprises a monitoring system including a submersible probe which is positioned in the batch solution to provide data to a controller. Once a desired level of nucleic acid compaction is reached, as detected by the monitoring system, the controller stops the dispensing and mixing agitating systems.

Abstract: A method is described for expressing biologically functional proteins without host cell toxicity. This method takes advantage of the surprising replication activating ability of the 107/402-T antigen. The invention also provides expression vectors, human cells, and fusion proteins for practicing the method.

Abstract: A pull-out keyboard tray. The pull-out keyboard tray includes an adjustable keyboard platform, removable storage compartments, and keyboard wiring protrusions. The keyboard is slightly mounted on drawer slides having multiple position detents, and is adapted for mounting under a desk top surface and the like. The keyboard further includes a removable palmrest, as well as slide shrouds for shrouding the drawer slides.

Abstract: A pull-out keyboard tray. The pull-out keyboard tray includes an adjustable keyboard platform, removable storage compartments, and keyboard wiring protrusions. The keyboard is slightly mounted on drawer slides having multiple position detents, and is adapted for mounting under a desk top surface and the like. The keyboard further includes a removable palmrest, as well as slide shrouds for shrouding the drawer slides.

Abstract: Counterions of polycations used to compact nucleic acids profoundly affect shape and stability of particles formed. Shape is associated with differential serum nuclease resistance and colloidal stability. A surrogate for determining such properties that is easy to measure is the turbidity parameter. Shape also affects the suitability and efficacy of compacted nucleic acid complexes for transfecting cells by various routes into a mammalian body.

Abstract: Novel methods are described for expression of biologically functional therapeutic and other proteins without host cell toxicity. The methods take advantage of the surprising replication activating ability of the 107/402-T antigen. The invention also provides fusion proteins, expression vectors, and mammalian cells, for practicing the methods.

Abstract: Episomal plasmids containing a papovavirus origin of replication and a papovavirus large T antigen mutant form are shown to replicate episomally in human cells, and yield levels of gene expression proportional to their episomal copy number. In conjunction with liposomal or receptor-mediated delivery systems, papovavirus-derived episomal plasmids provide an alternative vector for gene therapy, particularly when utilizing strategies requiring high levels of gene expression.

Abstract: An automated nucleic acid compaction device for analyzing and compacting a nucleic acid complex into unimolecular nucleic acid particles. Broadly, the device includes a container support and agitation system; a measuring and testing system; and a dispensing system; all controlled by a control system. The control system controls the support and agitation system and the dispensing system based either on a predetermined formulation or by analysis of feedback data provided by the measuring and testing system. In a preferred embodiment, the device is a real-time measuring and mixing instrument operating in a closed loop system. The preferred embodiment also comprises a monitoring system including a submersible probe which is positioned in the batch solution to provide data to a controller. Once a desired level of nucleic acid compaction is reached, as detected by the monitoring system, the controller stops the dispensing and mixing agitating systems.