Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
|---|
| relative hypoglycemia and hyperinsulinemia in mice with heterozygous lipoprotein lipase (lpl) deficiency. islet lpl regulates insulin secretion. | lipoprotein lipase (lpl) provides tissues with fatty acids, which have complex effects on glucose utilization and insulin secretion. to determine if lpl has direct effects on glucose metabolism, we studied mice with heterozygous lpl deficiency (lpl+/-). lpl+/- mice had mean fasting glucose values that were up to 39 mg/dl lower than lpl+/+ littermates. despite having lower glucose levels, lpl+/- mice had fasting insulin levels that were twice those of +/+ mice. hyperinsulinemic clamp experiments ... | 1999 | 10488074 |
| immune responses to adenovirus and adeno-associated virus in humans. | vectors based on human adenovirus (ad) and adeno-associated virus (aav) are being evaluated for human gene therapy. the response of the host to the vector, in terms of antigen-specific immunity, will play a substantial role in clinical outcome. we have surveyed cohorts of normal subjects and cystic fibrosis patients for pre-existing immunity to these viruses, caused by naturally acquired infections. a number of humoral and cellular assays to adenovirus serotype 5 (ad5) and adeno-associated virus ... | 1999 | 10490767 |
| induction of apoptosis by cadmium and the adeno-associated virus rep proteins. | the parvoviruses exert antiproliferative effects on transformed cells in culture. the development of cell lines that inducibly express the parvovirus nonstructural proteins have implicated these proteins in the limitation of cell growth. to study the host cell interactions of the nonstructural proteins we have developed a human 293 cell line that expresses the adeno-associated virus (aav) rep gene upon induction with heavy metal salts. when induced with both zn(2+) and cd(2+), rep protein expres ... | 1999 | 10497113 |
| persistent, therapeutically relevant levels of human granulocyte colony-stimulating factor in mice after systemic delivery of adeno-associated virus vectors. | adeno-associated virus (aav) vectors have been shown to preferentially transduce hepatocytes after systemic administration in adult mice and to provide long-term expression of introduced genes. one application of this technology would be for the production of granulocyte colony-stimulating factor (g-csf), which increases mature neutrophil numbers in humans and in animals, and has therapeutic effects in disorders featuring chronic neutropenia, including cyclic, severe congenital, and idiopathic n ... | 1999 | 10498245 |
| chromosomal latency and expression at map unit 96 of a wild-type plus adeno-associated virus (aav)/neo vector and identification of p81, a new aav transcriptional promoter. | human adeno-associated virus (aav) is ubiquitous and known to establish latency by chromosomal integration. we have constructed a wild-type plus aav vector, ins96-0.9neo, containing the neomycin resistance gene open reading frame (neo orf) of 960 bases in length at map unit 96 of the virus. ins96-0.9neo was constructed in an unconventional manner in that the neo orf lacked a dedicated heterologous promoter. in this study, this wild-type plus aav vector was to used to test aav's packaging capacit ... | 1999 | 10774553 |
| maxillary sinusitis as a surrogate model for cf gene therapy clinical trials in patients with antrostomies. | assessing the biological activity and clinical efficacy of gene therapy is critically important in cystic fibrosis (cf). it is widely accepted that clinical testing using surrogate markers including pulmonary function will be useful in assessing clinical efficacy. one problem with pulmonary surrogate markers of cf disease is the large number of patients and length of time required to demonstrate clinical efficacy. an alternative to pulmonary testing of new cf treatments is use of the maxillary s ... | 1999 | 10738581 |
| viral gene delivery. | experimental studies of viral gene delivery generally support the principle that virus-mediated gene transfer is indeed possible. however, the field of gene therapy has not yet been realised as a practicable clinical intervention. the delay in translation of laboratory work to clinical utility largely reflects the inability of gene delivery vectors to convey adequate genetic material to a desired location, with adequate durability and low enough toxicity to be effective. current studies of viral ... | 1999 | 11139845 |
| transplacental drug delivery: gene and virus delivery to the trophoblast. | the development of successful strategies for delivering genes to the placenta may provide new opportunities for modifying trophoblast function in order to learn more about trophoblast physiology and to offer novel therapeutic options for complications of pregnancy that result from placental dysfunction. replication-deficient recombinant viral vectors are useful vehicles for introducing genes into cells in vitro and in vivo. recombinant adenovirus and herpes simplex virus vectors are unable to ef ... | 1999 | 10837747 |
| detection of adeno-associated virus type 2 in patients with viral infection. | 1999 | 10816616 | |
| oligodendrocyte-specific gene expression in mouse brain: use of a myelin-forming cell type-specific promoter in an adeno-associated virus. | to explore the feasibility of cell type-specific gene expression in oligodendrocytes as a possible therapeutic approach for demyelinating diseases, the cell specificity, tissue specificity, and duration of gene expression were investigated using recombinant adeno-associated viral vectors (raav) carrying a green fluorescence protein (gfp) gene. recombinant aav vectors carrying either the myelin basic protein (mbp) promoter (raav-mbp-gfp) or the cytomegalovirus (cmv) immediate early promoter (raav ... | 1999 | 10723060 |
| gene therapy in the inner ear. mechanisms and clinical implications. | the application of gene therapy to the inner ear is an emerging field of study. most studies report the expression of marker genes (e.g., galactosidase) within the tissues of the cochlea. the first biologic response of an inner ear tissue (i.e., auditory neurons) to transduction by a gene therapy vector expressing a therapeutic gene (a herpes amplicon vector containing a bdnf gene) was observed in spiral explants obtained from early postnatal rat cochleae. this study was important because it dem ... | 1999 | 10842605 |
| gene transfer into the cns using recombinant adeno-associated virus: analysis of vector dna forms resulting in sustained expression. | recombinant adeno-associated virus (raav) vectors have shown significant promise as vehicles for in vivo gene transfer, particularly for transduction of organs composed primarily of non-dividing cells (i.e., muscle, cns, and liver). however, the mechanistic basis for this desirable property remains unclear. to investigate the fate of raav genomes in mouse brain, we stereotactically injected an raav vector carrying the e. coli lacz gene into the caudate of balb/c mice and demonstrate efficient tr ... | 1999 | 10682906 |
| safety and biological efficacy of an adeno-associated virus vector-cystic fibrosis transmembrane regulator (aav-cftr) in the cystic fibrosis maxillary sinus. | the host immune response and low vector efficiency have been key impediments to effective cystic fibrosis transmembrane regulator (cftr) gene transfer for cystic fibrosis (cf). an adeno-associated virus vector (aav-cftr) was used in a phase i dose-escalation study to transfer cftr cdna into respiratory epithelial cells of the maxillary sinus of 10 cf patients. | 1999 | 10890777 |
| recombinant adeno-associated virus-based vectors provide short-term rather than long-term transduction of primitive hematopoietic stem cells. | bone marrow stem cells collected from b6-gpi-1a mice pretreated with 5-fluorouracil were incubated for 2 h at 37 degrees c in the presence of the recombinant adenovirus-associated virus-based vector (raav) ssv9. as measured in vitro immediately following transduction, ssv9 was found to be effective in transducing the primitive cobble-stone-area-forming cell (cafc)-35 subset (60% transduction efficiency). however, this did not predict long-term expression as the presence of the transgene could no ... | 1999 | 10195572 |
| transfer of activation-dependent gene expression into t cell lines by recombinant adeno-associated virus. | we examined the ability of recombinant adeno-associated virus (raav) to transfer regulated gene expression into t cell lines. an aav-based vector containing the neomycin resistance gene and expressing the firefly luciferase (luc) gene under the regulatory control of the interleukin 2 promoter (paav-luc) was generated and adenovirus-free raav (raav-luc) was produced from this vector. transfection of paav-luc into the human t cell line jurkat resulted in luciferase expression while infection of ju ... | 1999 | 10435102 |
| adeno-associated virus (aav) type 5 rep protein cleaves a unique terminal resolution site compared with other aav serotypes. | adeno-associated virus (aav) replication depends on two viral components for replication: the aav nonstructural proteins (rep) in trans, and inverted terminal repeat (itr) sequences in cis. aav type 5 (aav5) is a distinct virus compared to the other cloned aav serotypes. whereas the rep proteins and itrs of other serotypes are interchangeable and can be used to produce recombinant viral particles of a different serotype, aav5 rep proteins cannot cross-complement in the packaging of a genome with ... | 1999 | 10196327 |
| functional expression of exogenous proteins in mammalian sensory hair cells infected with adenoviral vectors. | to understand the function of specific proteins in sensory hair cells, it is necessary to add or inactivate those proteins in a system where their physiological effects can be studied. unfortunately, the usefulness of heterologous expression systems for the study of many hair cell proteins is limited by the inherent difficulty of reconstituting the hair cell's exquisite cytoarchitecture. expression of exogenous proteins within hair cells themselves may provide an alternative approach. because re ... | 1999 | 10200223 |
| nuclear transport of the major capsid protein is essential for adeno-associated virus capsid formation. | adeno-associated virus capsids are composed of three proteins, vp1, vp2, and vp3. although vp1 is necessary for viral infection, it is not essential for capsid formation. the other capsid proteins, vp2 and vp3, are sufficient for capsid formation, but the functional roles of each protein are still not well understood. by analyzing a series of deletion mutants of vp2, we identified a region necessary for nuclear transfer of vp2 and found that the efficiency of nuclear localization of the capsid p ... | 1999 | 10438891 |
| adeno-associated virus and development of cervical neoplasia. | evidence from several sources has suggested that adeno-associated virus (aav) infection might protect against cervical cancer, in part, by interfering with human papillomavirus (hpv)-induced tumorigenesis. detection of aav type 2 (aav-2) dna in cervical tissues has been reported. however, there have been few in vivo studies of women with cervical hpv infection or neoplasia, and these have reported inconsistent results. therefore, we used polymerase chain reaction (pcr) assays targeted to the aav ... | 1999 | 10440809 |
| stable restoration of the sarcoglycan complex in dystrophic muscle perfused with histamine and a recombinant adeno-associated viral vector. | limb-girdle muscular dystrophies 2c-f represent a family of autosomal recessive diseases caused by defects in sarcoglycan genes. the cardiomyopathic hamster is a naturally occurring model for limb-girdle muscular dystrophy caused by a primary deficiency in delta-sarcoglycan. we show here that acute sarcolemmal disruption occurs in this animal model during forceful muscle contraction. a recombinant adeno-associated virus vector encoding human delta-sarcoglycan conferred efficient and stable genet ... | 1999 | 10202936 |
| enhanced sensitivity of small cell lung cancer cell lines to cisplatin and etoposide after infection with adeno-associated virus type 2. | in previous studies we have reported the sensitisation of human tumour cells to gamma irradiation and chemotherapeutic drugs upon infection with the human non-pathogenic adeno-associated virus type 2 (aav-2) in vitro and in vivo. treatment of small cell lung cancer (sclc) is consistently hampered by relapses due to the selection of chemotherapy-resistant cell clones. hence, we were interested to test whether selection of chemotherapy-resistant sclc cells might be reduced or even prevented if che ... | 1999 | 10211097 |
| hepatic gene therapy using adeno-associated virus vectors. | recombinant adeno-associated virus vectors have been shown to safely transduce a number of tissues in preclinical animal studies. the level of gene transfer is sufficient to successfully treat a large number of medical disorders. moreover, the long-term persistence of the vector sequences in animals makes it likely that this vector will be useful for genetic diseases requiring life-long therapy. this review outlines the biological principles of the vector, as well as its advantages and current l ... | 1999 | 10349684 |
| interaction of adeno-associated virus rep78 with p53: implications in growth inhibition. | adeno-associated virus (aav) is a nonpathogenic, single-stranded dna virus belonging to the parvoviridae family. onco-suppressive properties of aav against adenovirus, a dna tumor virus, have been well documented. rep78, a major regulatory protein of aav, is believed to be responsible for its antioncogenic properties. most dna tumor viruses disturb the cell cycle pathways by essentially abrogating the functions of p53. here we present evidence that aav acts as an antiproliferative agent against ... | 1999 | 10446967 |
| gene transfer into the mammalian inner ear using hsv-1 and vaccinia virus vectors. | the introduction of foreign genes into cells has become an effective means of achieving intracellular expression of foreign proteins, both for therapeutic purposes and for experimental manipulation. gene delivery to the nervous system has been extensively studied, primarily using viral vectors. however, to date less work has focused on gene delivery to the inner ear, and existing studies have primarily used adenovirus and adeno-associated virus. using two recombinant viral vectors, herpes simple ... | 1999 | 10452370 |
| long-term actions of vector-derived nerve growth factor or brain-derived neurotrophic factor on choline acetyltransferase and trk receptor levels in the adult rat basal forebrain. | trophic factor gene therapy may provide a rational treatment strategy for neurodegenerative disease. recombinant adeno-associated virus vectors, incorporating a neuron-specific promoter driving bicistronic expression of green fluorescent protein and either nerve growth factor or brain-derived neurotrophic factor, transduced 10,000-15,000 neurons in the medial septum for periods of at least six months. both cholinergic and non-cholinergic neurons expressed green fluorescent protein. nerve growth ... | 1999 | 10218782 |
| highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses. | gene transfer vectors based on the replication-defective human parvovirus, adeno-associated virus type 2 (aav-2), are viable candidates for in vivo and ex vivo human use. however, widespread testing of aav vectors has been limited by difficulties in generating pure, high-titer vector stocks that are fully characterized. to address these issues, we have developed a single-step purification scheme using heparin affinity chromatography. recovery from the crude lysate starting material exceeds 70%, ... | 1999 | 10223736 |
| adeno-associated virus 2 co-receptors? | 1999 | 10229209 | |
| position-independent human beta-globin gene expression mediated by a recombinant adeno-associated virus vector carrying the chicken beta-globin insulator. | the position-independent expression of transgenes in target cells is an essential subject for determining effective gene therapies. the chicken beta-globin insulator blocks the effects of regulatory sequences on transcriptional units at differential domains. we prepared a recombinant adeno-associated virus (raav) containing various combinations of the dnase i-hypersensitive site 2 (hs2), 3 (hs3), and 4 (hs4) core elements from the human beta-globin locus control region (lcr), the human beta-glob ... | 1999 | 10319578 |
| raav vector-mediated sarcogylcan gene transfer in a hamster model for limb girdle muscular dystrophy. | the limb girdle muscular dystrophies (lgmd) are a genetically and phenotypically heterogeneous group of degenerative neuromuscular diseases. a subset of the genetically recessive forms of lgmd are caused by mutations in the four muscle sarcoglycan genes (alpha, beta, gamma and delta). the coding sequences of all known sarcoglycan genes are smaller than 2 kb, and thus can be readily packaged in recombinant adeno-associated virus (raav) vectors. previously, we have demonstrated highly efficient an ... | 1999 | 10341878 |
| ca2+ and phorbol ester effect on the mast cell phosphoprotein induced by cromolyn. | several phosphoproteins are involved in stimulus-secretion coupling. the beta and gamma subunits of immunoglobulin e binding protein (fc epsilonri) and three other protein bands get phosphorylated during stimulation of mast cell secretion. these additional proteins of 42, 59 and 68 kda are also phosphorylated when secretion is stimulated by compound 48/80 (c48/80). a 78 kda band, however, is phosphorylated as secretion wanes after stimulation with c48/80 and by the anti-allergic drug disodium cr ... | 1999 | 10357262 |
| efficient production of adeno-associated virus vectors using split-type helper plasmids. | adeno-associated virus (aav) vectors are potentially useful vehicles for the delivery of therapeutic genes into human cells. to determine the optimal expression pattern of aav proteins (rep78, rep68, rep52, rep40, and cap proteins) for packaging the recombinant aav genome, helper plasmids were split into two portions. in this study, two sets of split-type helper plasmids were prepared; i.e., 1) a rep expression plasmid (prep) and cap expression plasmid (pcap), and 2) a large rep expression plasm ... | 1999 | 10363588 |
| cellular redox state alters recombinant adeno-associated virus transduction through tyrosine phosphatase pathways. | several types of environmental damage including uv, hydroxyurea and ionizing irradiation have been shown to augment raav transduction. current hypotheses suggest that these environmental stimuli lead to the enhanced production and/or activation of cellular factors important in the conversion of single-stranded dna genomes to expressible forms. however, the mechanisms of action are currently unknown. we hypothesized that reactive oxygen intermediates (roi) may play a common role in the augmentati ... | 1999 | 10467367 |
| isolation of recombinant adeno-associated virus vector-cellular dna junctions from mouse liver. | recombinant adeno-associated virus (raav) vectors allow for sustained expression of transgene products from mouse liver following a single portal vein administration. here a raav vector expressing human coagulation factor f.ix (hf.ix), aav-ef1alpha-f.ix (hf.ix expression was controlled by the human elongation factor 1alpha [ef1alpha] enhancer-promoter) was injected into mice via the portal vein or tail vein, or directly into the liver parenchyma, and the forms of raav vector dna extracted from t ... | 1999 | 10364291 |
| genetic capsid modifications allow efficient re-targeting of adeno-associated virus type 2. | the human parvovirus adeno-associated virus type 2 (aav2) has many features that make it attractive as a vector for gene therapy. however, the broad host range of aav2 might represent a limitation for some applications in vivo, because recombinant aav vector (raav)-mediated gene transfer would not be specific for the tissue of interest. this host range is determined by the binding of the aav2 capsid to specific cellular receptors and/or co-receptors. the tropism of aav2 might be changed by genet ... | 1999 | 10470084 |
| transduction of well-differentiated airway epithelium by recombinant adeno-associated virus is limited by vector entry. | the limitations of adeno-associated virus (aav)-mediated vectors for lung-directed gene transfer were investigated by using differentiated human respiratory epithelium in air-liquid interface cultures. transduction efficiency was high in undifferentiated cells and was enhanced in well-differentiated cells after basolateral application of the vector or after apical application following disruption of tight junctions or pretreatment of the cultures with glycosidases. these results indicate that tr ... | 1999 | 10364362 |
| suppression of tumorigenicity in cervical carcinoma hela cells by an episomal form of adeno-associated virus. | to compare the oncosuppressive activity of integrated and episomal adeno-associated virus (aav), two epstein-barr virus (ebv) derived episomal form vectors, p205 and p220, were used to generate plasmids containing episomal aav. hela cells transfected with p205, p220, and the plasmids with forward (pa205-1, pa220-1) and reverse (pa205-1, pa220-2) orientation of inserted aav dna were designated h205, h220, ha205-1, ha220-1, ha205-1, and ha220-2, respectively. the respective average copy numbers of ... | 1999 | 10375600 |
| factors affecting the terminal resolution site endonuclease, helicase, and atpase activities of adeno-associated virus type 2 rep proteins. | the rep68 and rep78 proteins (rep68/78) of adeno-associated virus type 2 (aav) are critical for aav replication and site-specific integration. they bind specifically to the aav inverted terminal repeats (itrs) and possess atpase, helicase, and strand-specific/site-specific endonuclease activities. in the present study, we further characterized the aav rep68/78 helicase, atpase, and endonuclease activities by using a maltose binding protein-rep68 fusion (mbp-rep68delta) produced in escherichia co ... | 1999 | 10482574 |
| generation of aberrant sprouting in the adult rat brain by gap-43 somatic gene transfer. | the expression of gap-43 was modulated genetically in the adult rat nigrostriatal or septohippocampal pathway using recombinant adeno-associated virus (raav) vectors incorporating the neuron specific enolase (nse) promoter and either a rat gap-43 cdna or the corresponding antisense sequence. bicistronic expression of green fluorescent protein (gfp) enabled us to evaluate transduced neurons selectively. single injections of raav into the substantia nigra pars compacta (snc) transduced both dopami ... | 1999 | 10375659 |
| [effect of mechanical damage on ex vivo dna virus vector-mediated gene transduction in epithelial cells of murine trachea]. | the mechanism of adenovirus (ad) and adeno-associated virus (aav) vector-mediated gene transduction in murine tracheae has not been fully understood. excised tracheae from mice were exposed to either ad vector (ad-cmv-lacz) or aav vector (aav-cmv-lacz) for 1 hour. lacz gene expression in tracheal epithelial cells was detected by x-gal staining. only patch distributions of lacz expressing cells were observed. the percentage of lacz expressing cells to total cells was less than 1% with either vect ... | 1999 | 10390962 |
| long-term restoration of striatal l-aromatic amino acid decarboxylase activity using recombinant adeno-associated viral vector gene transfer in a rodent model of parkinson's disease. | as a potential treatment for parkinson's disease, viral vector-mediated over-expression of striatal l-aromatic amino acid decarboxylase was tested in an attempt to facilitate the production of therapeutic levels of dopamine after peripheral l-dihydroxyphenylalanine administration. the results of microdialysis and enzyme activity assays indicate that striatal decarboxylation of peripherally administered l-dihydroxyphenylalanine was enhanced by recombinant adeno-associated virus-mediated gene tran ... | 1999 | 10392841 |
| persistent transgene product in retina, optic nerve and brain after intraocular injection of raav. | recombinant adeno-associated virus (raav) is a promising vector for retinal application as it transduces photoreceptors and retinal pigment epithelium cells efficiently and in a stable fashion. because raav also transduces retinal ganglion cells, we reasoned that ocular application of raav might result in delivery of transgenic protein to the cns. here we describe high levels of green fluorescent protein (gfp) persisting at least 6 months in optic nerves and brains of mice and dogs after intravi ... | 1999 | 10396623 |
| recombinant adeno-associated virus (aav) drives constitutive production of glutamate decarboxylase in neural cell lines. | many neurological disorders result directly or indirectly from the loss of inhibitory function. engineering the production of gaba, an inhibitory neurotransmitter, may therefore be able at least partly to restore the lost inhibition seen in epilepsy, parkinson's disease, or huntington's disease. in this article, we describe a set of recombinant adeno-associated viruses (aavs) that can deliver cdnas encoding the gaba-producing enzyme, glutamate decarboxylase (gad), directly into neural cells. we ... | 1999 | 10397644 |
| long-term regulated expression of growth hormone in mice after intramuscular gene transfer. | effective delivery of secreted proteins by gene therapy will require a vector that directs stable delivery of a transgene and a regulatory system that permits pharmacologic control over the level and kinetics of therapeutic protein expression. we previously described a regulatory system that enables transcription of a target gene to be controlled by rapamycin, an orally bioavailable drug. here we demonstrate in vivo regulation of gene expression after intramuscular injection of two separate aden ... | 1999 | 10411931 |
| adeno-associated virus vectors and hematology. | 1999 | 10419876 | |
| adeno-associated viral vectors for gene transfer and gene therapy. | adeno-associated virus (aav) is a defective, non-pathogenic human parvovirus that depends for growth on coinfection with a helper adenovirus or herpes virus. recombinant adeno-associated viruses (raavs) have attracted considerable interest as vectors for gene therapy. in contrast to other gene delivery systems, raavs lack all viral genes and show long-term gene expression in vivo without immune response or toxicity. over the past few years, many applications of raavs as therapeutic agents have d ... | 1999 | 10430026 |
| efficient gene transfer into human keratinocytes with recombinant adeno-associated virus vectors. | gene transfer into the skin is a promising approach to treat inherited or acquired dermatological diseases and systemic monogenic deficiencies. for this purpose, the efficient and sustained gene delivery into keratinocytes is of critical importance. recombinant adeno-associated virus (raav) vectors hold the potential to achieve a long-term gene transfer into various human organs. in order to evaluate this potential for skin gene therapy, human keratinocytes were transduced in vitro with raav vec ... | 1999 | 10435093 |
| submucosal gland development in the airway is controlled by lymphoid enhancer binding factor 1 (lef1). | previous studies have demonstrated that transcription of the lymphoid enhancer binding factor 1 (lef1) gene is upregulated in submucosal gland progenitor cells just prior to gland bud formation in the developing ferret trachea. in the current report, several animal models were utilized to functionally investigate the role of lef1 in initiating and supporting gland development in the airway. studies on lef1-deficient mice and antisense oligonucleotides in a ferret xenograft model demonstrate that ... | 1999 | 10498680 |
| production of recombinant adeno-associated virus vectors using a packaging cell line and a hybrid recombinant adenovirus. | recombinant adeno-associated virus (raav) vectors are under consideration for a wide variety of gene therapy applications. one of the limitations of the raav vector system has been the difficulty in producing the vector in sufficient quantity for adequate preclinical and clinical evaluation. a common method for vector production involves large-scale transient transfection of multiple plasmids into cultured cells. because this approach might not be feasible for clinical scale manufacturing, we ha ... | 1999 | 10435114 |
| high-fidelity correction of mutations at multiple chromosomal positions by adeno-associated virus vectors. | the gene targeting techniques used to modify chromosomes in mouse embryonic stem cells have had limited success with many other cell types, especially normal primary cells with restricted growth capacity outside the organism. this is due in large part to the technical problems and/or inefficiency of conventional dna transfer methods, as well as the low rates of homologous recombination obtained in unselected cell populations. we recently described an alternative approach in which adeno-associate ... | 1999 | 10438827 |
| application of gene therapy to acute inflammatory diseases. | the application of gene therapy to acute inflammation has not received as much research attention as has the treatment of genetically-based diseases, cancer, and viral infections. however, gene therapy as a drug delivery system offers several theoretical and practical advantages over current protein delivery systems. these include the ability to target therapies to individual tissues or cell types, to locally produce proteins that can act intracellularly or in an autocrine, juxtacrine, or paracr ... | 1999 | 10446888 |
| enhanced expression of transgenes from adeno-associated virus vectors with the woodchuck hepatitis virus posttranscriptional regulatory element: implications for gene therapy. | the woodchuck hepatitis virus posttranscriptional regulatory element (wpre) evolved to stimulate the expression of intronless viral messages. to determine whether this ability to enhance expression could be useful in nonviral and heterologous viral gene delivery systems, we analyzed the ability of the wpre to elevate the expression of a cdna encoding the green fluorescent protein (gfp) in these contexts. we find that the wpre can stimulate the expression of gfp when the gene is delivered by tran ... | 1999 | 10515449 |
| purification of recombinant adeno-associated virus by iodixanol gradient ultracentrifugation allows rapid and reproducible preparation of vector stocks for gene transfer in the nervous system. | recombinant adeno-associated virus (raav) vectors have become attractive tools for in vivo gene transfer. the production and purification of high-titer raav vector stocks for experimental and therapeutic gene transfer continue to undergo improvement. standard raav vector purification protocols include the purification of the vector by cesium chloride (cscl)-density gradient centrifugation followed by extensive desalination via dialysis against a physiological buffer for in vivo use. these proced ... | 1999 | 10446928 |
| adeno-associated virus as a gene delivery vector for liver cells. | 1999 | 10453960 | |
| recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. | conventional methods for raav purification that are based on cesium chloride ultracentrifugation have often produced vector preparations of variable quality and resulted in significant loss of particle infectivity. we report here several novel purification strategies that involve the use of non-ionic iodixanol gradients followed by ion exchange or heparin affinity chromatography by either conventional or hplc columns. these methods result in more than 50% recovery of raav from a crude lysate and ... | 1999 | 10455399 |
| charge-to-alanine mutagenesis of the adeno-associated virus type 2 rep78/68 proteins yields temperature-sensitive and magnesium-dependent variants. | the adeno-associated virus type 2 (aav) replication (rep) proteins rep78 and 68 (rep78/68) exhibit a number of biochemical activities required for aav replication, including specific binding to a 22-bp region of the terminal repeat, site-specific endonuclease activity, and helicase activity. individual and clusters of charged amino acids were converted to alanines in an effort to generate a collection of conditionally defective rep78/68 proteins. rep78 variants were expressed in human 293 cells ... | 1999 | 10516052 |
| repeated delivery of adeno-associated virus vectors to the rabbit airway. | efficient local expression from recombinant adeno-associated virus (raav)-cystic fibrosis (cf) transmembrane conductance regulator (cftr) vectors has been observed in the airways of rabbits and monkeys for up to 6 months following a single bronchoscopic delivery. however, it is likely that repeated administrations of raav vectors will be necessary for sustained correction of the cf defect in the airways. the current study was designed to test the feasibility of repeated airway delivery of raav v ... | 1999 | 10516053 |
| concatamerization of adeno-associated virus circular genomes occurs through intermolecular recombination. | long-term recombinant aav (raav) transgene expression in muscle has been associated with the molecular conversion of single-stranded raav genomes to high-molecular-weight head-to-tail circular concatamers. however, the mechanisms by which these large multimeric concatamers form remain to be defined. to this end, we tested whether concatamerization of raav circular intermediates occurs through intra- or intermolecular mechanisms of amplification. coinfection of the tibialis muscle of mice with ra ... | 1999 | 10516055 |
| persistent expression of canine factor ix in hemophilia b canines. | we previously demonstrated that direct intramuscular injection of recombinant adeno-associated virus (raav) carrying the human fix (hfix) cdna can safely be administered to hemophilic b canines and express human factor ix protein; however, the functional activity of the hfix protein could not be assessed due to anti-human fix antibody (inhibitor) formation. to test the therapeutic efficacy of raav in hemophilic dogs, raav type 2 (raav2) carrying canine fix (cfix) cdna was injected into the skele ... | 1999 | 10516718 |
| binding of the human papillomavirus type 16 p97 promoter by the adeno-associated virus rep78 major regulatory protein correlates with inhibition. | human papillomavirus type 16 (hpv-16) infection is positively associated with cervical cancer, whereas adeno-associated virus (aav) infection is negatively associated with this same cancer. in earlier studies these two virus types have been shown to directly interact, with aav inhibiting or enhancing papillomavirus functions depending upon the specific circumstances. one defined interaction between these two viruses is the ability of the aav rep78 major regulatory protein to inhibit gene express ... | 1999 | 10531369 |
| elimination of lysosomal storage in brains of mps vii mice treated by intrathecal administration of an adeno-associated virus vector. | mucopolysaccharidosis type vii (mps vii) is an inherited lysosomal storage disease caused by insufficient beta-glucuronidase (gus). to provide gene therapy in a mutant mouse model of this disease, we have used a recombinant adeno-associated virus (raav) vector to deliver gus cdna to a variety of tissues. although intravenous administration of vector produced therapeutic levels of gus in the liver, delivery to the brain was inadequate. to improve delivery to the brain intrathecal injection of the ... | 1999 | 10455422 |
| recombinant aav-2 harboring gfp-antisense/ribozyme fusion sequences monitor transduction, gene expression, and show anti-hiv-1 efficacy. | vector-mediated delivery of potentially antivirally active genes is a key step in somatic gene therapy including therapeutic approaches against aids. a crucial technical prerequisite is to monitor dna transfer into target cells. here, we describe recombinant infectious particles derived from the adeno-associated virus type 2 (aav-2) that are suitable to deliver effective hiv-1-directed antisense and ribozyme genes into target cells. to monitor transduction, we designed and tested a number of fus ... | 1999 | 10455431 |
| stable transgene expression in rod photoreceptors after recombinant adeno-associated virus-mediated gene transfer to monkey retina. | recombinant adeno-associated virus (raav) is a promising vector for therapy of retinal degenerative diseases. we evaluated the efficiency, cellular specificity, and safety of retinal cell transduction in nonhuman primates after subretinal delivery of an raav carrying a cdna encoding green fluorescent protein (egfp), raav. cmv.egfp. the treatment results in efficient and stable egfp expression lasting >1 year. transgene expression in the neural retina is limited exclusively to rod photoreceptors. ... | 1999 | 10449795 |
| gene transfer to neurons with herpes simplex virus/adeno-associated virus hybrid vectors. | 1999 | 10543609 | |
| progress in adeno-associated virus type 2 vector production: promises and prospects for clinical use. | vectors derived from the human parvovirus aav-2 (adeno-associated virus type 2) are among the most promising gene delivery vehicles currently being developed. these vectors are not only capable of transducing a large variety of human cell types in vitro and in vivo, but in immunocompetent animal models can establish long-term gene expression without being pathogenic to the recipient. however, a limitation of this vector system with respect to its clinical application has long been the laborious ... | 1999 | 10543610 |
| gene transfer to the nigrostriatal system by hybrid herpes simplex virus/adeno-associated virus amplicon vectors. | to improve gene transfer to cns neurons, critical elements of herpes simplex virus 1 (hsv-1) amplicons and recombinant adeno-associated virus (aav) vectors were combined to construct a hybrid amplicon vector, and then packaged via a helper virus-free system. we tested the hsv/aav hybrid amplicon vectors for transduction efficiency and stability of transgene expression (green fluorescent protein) in primary neuronal cultures from rat fetal ventral mesencephalon, in comparison with traditional hsv ... | 1999 | 10543613 |
| high-titer recombinant adeno-associated virus production from replicating amplicons and herpes vectors deleted for glycoprotein h. | production of high-titer raav is essential for in vivo clinical application. one limiting factor may be the failure of existing systems to replicate the packaging genome in such a way that expression of rep and cap proteins is coordinately amplified. disc-hsv (disabled single-cycle virus) is a genetically modified herpes simplex virus (hsv) that by deletion of glycoprotein h (gh) is infectious only if propagated in a complementing cell line. in this study, we have used disc-hsv as a helper for r ... | 1999 | 10543617 |
| rep-mediated nicking of the adeno-associated virus origin requires two biochemical activities, dna helicase activity and transesterification. | the single-stranded adeno-associated virus (aav) genome is flanked by terminal hairpinned origins of dna replication (terminal repeats [trs]) that are nicked at the terminal resolution site (trs) by the aav rep protein in an atp-dependent, site-specific manner. here we determine the minimal trs sequence necessary for rep cleavage, 3'-ccggt/tg-5', and show that this 7-base core sequence is required only on the nicked strand. we also identify a potential stem-loop structure at the trs. interesting ... | 1999 | 10516041 |
| superior gene transfer into solid tumour cells than into human mobilised peripheral blood progenitor cells using helpervirus-free adeno-associated viral vector stocks. | autologous peripheral blood progenitor cell (pbpc) grafts can be contaminated with tumour cells that potentially give rise to relapse following myeloablative therapy and pbpc transplantation. adeno-associated virus (aav)-based vectors produced by a new adenovirus-free technique are a gene delivery system which may be applicable for tumour cell purging. to test for the host range of these vectors, solid tumours of clinical relevance and normal cd34+ pbpc were selected as target cells for an aav-v ... | 1999 | 10533460 |
| dynamin is required for recombinant adeno-associated virus type 2 infection. | recombinant adeno-associated virus (raav) vectors for gene therapy of inherited disorders have demonstrated considerable potential for molecular medicine. recent identification of the viral receptor and coreceptors for aav type 2 (aav-2) has begun to explain why certain organs may demonstrate higher efficiencies of gene transfer with this vector. however, the mechanisms by which aav-2 enters cells remain unknown. in the present report, we have examined whether the endocytic pathways of raav-2 ar ... | 1999 | 10559355 |
| high-titer recombinant adeno-associated virus production utilizing a recombinant herpes simplex virus type i vector expressing aav-2 rep and cap. | recombinant adeno-associated virus type 2 (raav) vectors have recently been used to achieve long-term, high level transduction in vivo. further development of raav vectors for clinical use requires significant technological improvements in large-scale vector production. in order to facilitate the production of raav vectors, a recombinant herpes simplex virus type i vector (rhsv-1) which does not produce icp27, has been engineered to express the aav-2 rep and cap genes. the optimal dose of this v ... | 1999 | 10455400 |
| gene therapy vectors based on adeno-associated virus type 1. | the complete sequence of adeno-associated virus type 1 (aav-1) was defined. its genome of 4,718 nucleotides demonstrates high homology with those of other aav serotypes, including aav-6, which appears to have arisen from homologous recombination between aav-1 and aav-2. analysis of sera from nonhuman and human primates for neutralizing antibodies (nab) against aav-1 and aav-2 revealed the following. (i) nab to aav-1 are more common than nab to aav-2 in nonhuman primates, while the reverse is tru ... | 1999 | 10196295 |
| cloning and characterization of adeno-associated virus type 5. | adeno-associated virus type 5 (aav5) is distinct from other dependovirus serotypes based on dna hybridization and serological data. to better understand the biology of aav5, we have cloned and sequenced its genome and generated recombinant aav5 particles. the single-stranded dna genome is similar in length and genetic organization to that of aav2. the rep gene of aav5 is 67% homologous to aav2, with the majority of the changes occurring in the carboxyl and amino termini. this homology is much le ... | 1999 | 9882336 |
| biochemical characterization of adeno-associated virus rep68 dna helicase and atpase activities. | the adeno-associated virus (aav) nonstructural proteins rep68 and rep78 are site-specific dna binding proteins, atp-dependent site-specific endonucleases, helicases, and atpases. these biochemical activities are required for viral dna replication and control of viral gene expression. in this study, we characterized the biochemical properties of the helicase and atpase activities of homogeneously pure rep68. the enzyme exists as a monomer in solution at the concentrations used in this study (<380 ... | 1999 | 9882364 |
| antisense inhibition of at1 receptor in vascular smooth muscle cells using adeno-associated virus-based vector. | vascular smooth muscle cells (vsmcs) are the main peripheral target for vasoconstriction and growth-promoting activity of angiotensin ii (ang ii), acting through angiotensin type 1 receptors (at1-r). current antihypertension treatments include daily reductions in the effects of ang ii. to decrease an effect of ang ii in a prolonged fashion, we have developed an adeno-associated virus (aav) vector with antisense dna for at1-r. aav has many advantages over other viral vectors. aav is nonpathogenic ... | 1999 | 9931129 |
| gene therapy for cerebrovascular disease. | to review the principles of and the experimental and clinical results of gene therapy for cerebrovascular disease. | 1999 | 9932877 |
| structure of adeno-associated virus vector dna following transduction of the skeletal muscle. | the skeletal muscle provides a very permissive physiological environment for adeno-associated virus (aav) type 2-mediated gene transfer. we have studied the early steps leading to the establishment of permanent transgene expression, after injection of recombinant aav (raav) particles in the quadriceps muscle of mice. the animals received an raav encoding a secreted protein, murine erythropoietin (mepo), under the control of the human cytomegalovirus major immediate-early promoter and were sacrif ... | 1999 | 9971774 |
| development of animal models for adeno-associated virus site-specific integration. | the adeno-associated virus (aav) is unique in its ability to target viral dna integration to a defined region of human chromosome 19 (aavs1). since aavs1 sequences are not conserved in a rodent's genome, no animal model is currently available to study aav-mediated site-specific integration. we describe here the generation of transgenic rats and mice that carry the aavs1 3.5-kb dna fragment. to test the response of the transgenic animals to rep-mediated targeting, primary cultures of mouse fibrob ... | 1999 | 9971837 |
| neonatal intramuscular injection with recombinant adeno-associated virus results in prolonged beta-glucuronidase expression in situ and correction of liver pathology in mucopolysaccharidosis type vii mice. | for many metabolic diseases, early correction of the inherited deficiency is required to prevent long-term sequelae. we examined the ability of adeno-associated virus (aav) to mediate efficient gene transfer during the neonatal period in mice with the lysosomal storage disease mucopolysaccharidosis type vii (mps vii). quadriceps of newborn mps vii mice were injected with an aav vector containing human beta-glucuronidase (gusb) cdna. high-level intramuscular gusb expression was seen as early as 2 ... | 1999 | 10022533 |
| adeno-associated virus-mediated gene transfer to the brain: duration and modulation of expression. | adeno-associated virus (aav) is a promising vector for central nervous system (cns) gene transfer, but a number of issues must be addressed if aav is to be used for widespread delivery throughout the cns. our aim was to test the effect of dose, route of delivery, and hydroxyurea treatment on brain expression of beta-galactosidase activity after cerebral inoculation with an raav-lacz vector (raav-beta-gal). we also wished to test whether an immune response appeared against the vector and the tran ... | 1999 | 10022545 |
| a 110-kda nuclear shuttle protein, nucleolin, specifically binds to adeno-associated virus type 2 (aav-2) capsid. | a 110-kda protein was copurified with adeno-associated virus type 2 (aav-2) virions after cscl density gradient isopycnic centrifugation. amino acid sequence of peptides derived from this protein after tryptic digestion, monoclonal antibody production, and western blot analysis showed that the copurified protein was the major nucleolar phosphoprotein, human nucleolin. virus overlay assays demonstrated that aav-2 capsid specifically bound to the human nucleolin, and immunoprecipitation studies co ... | 1999 | 10329548 |
| recombinant adeno-associated virus-mediated expression of o6-alkylguanine-dna-alkyltransferase protects human epithelial and hematopoietic cells against chloroethylating agent toxicity. | recombinant adeno-associated virus (raav) encoding the human o6-alkylguanine-dna-alkyltransferase (hat) protein and a selectable marker (neo(r)) was used to transduce human cervical carcinoma (hela) cells and erythroleukemic (k562) cells and clones were selected using g418 (0.4 mg/ml). thirteen hela clones were isolated, 9 of which survived for 2-3 months before cell death ensued, presumably owing to the loss of g418 resistance. northern blot analysis of the remaining four clones, using a neo pr ... | 1999 | 10022554 |
| comparative characterization of rep proteins from the helper-dependent adeno-associated virus type 2 and the autonomous goose parvovirus. | adeno-associated viruses (aavs) are nonautonomous human parvoviruses in that they are dependent on helper functions supplied by other viruses or on genotoxic stimuli for conditions permissive for replication. in the absence of helper, aav type 2 enters latency by integration into a specific site on human chromosome 19. this feature of aav, in combination with a lack of pathogenicity, makes aav an attractive candidate vector for human gene therapy. goose parvovirus (gpv) is both autonomous and pa ... | 1999 | 10074142 |
| disease-inducible transgene expression from a recombinant adeno-associated virus vector in a rat arthritis model. | rheumatoid arthritis (ra) is a systemic autoimmune disease affecting 1% of the world's population, with significant morbidity and mortality. in this study, we investigated a recombinant adeno-associated virus (raav) vector for its potential application in ra gene therapy. raav encoding escherichia coli beta-galactosidase was injected into rat joints which had already been induced into acute arthritis after local lipopolysaccharide (lps) administration, and the efficiency of in vivo transduction ... | 1999 | 10074195 |
| formation of adeno-associated virus circular genomes is differentially regulated by adenovirus e4 orf6 and e2a gene expression. | a central feature of the adeno-associated virus (aav) latent life cycle is persistence in the form of both integrated and episomal genomes. however, the molecular processes associated with episomal long-term persistence of aav genomes are only poorly understood. to investigate these mechanisms, we have utilized a recombinant aav (raav) shuttle vector to identify circular aav intermediates from transduced hela cells and primary fibroblasts. the unique structural features exhibited by these transd ... | 1999 | 9847318 |
| analysis of the effects of charge cluster mutations in adeno-associated virus rep68 protein in vitro. | the rep78 and rep68 proteins of adeno-associated virus type 2 (aav) are multifunctional proteins which are required for viral replication, regulation of aav promoters, and preferential integration of the aav genome into a region of human chromosome 19. these proteins bind the hairpin structures formed by the aav inverted terminal repeat (itr) origins of replication, make site- and strand-specific endonuclease cuts within the aav itrs, and display nucleoside triphosphate-dependent helicase activi ... | 1999 | 9971790 |
| alphavbeta5 integrin: a co-receptor for adeno-associated virus type 2 infection. | understanding the primary steps of viral entry can have important implications for strategies to prevent infection of known viral pathogens as well as determining parameters for efficient gene delivery using viral vectors. recently, a two-step process for viral infection involving attachment of virus to a primary receptor (coxsackievirus adenovirus receptor and heparan sulfate proteoglycan) and subsequent mediation of virus entry by a co-receptor (alphav integrins and hvem) has been determined f ... | 1999 | 9883843 |
| efficient and stable transduction of cardiomyocytes after intramyocardial injection or intracoronary perfusion with recombinant adeno-associated virus vectors. | the delivery of recombinant genes to cardiomyocytes holds promise for the treatment of a variety of cardiovascular diseases. previous gene transfer approaches that used direct injection of plasmid dna or replication-defective adenovirus vectors have been limited by low transduction frequencies and transient transgene expression due to immune responses, respectively. in this report, we have tested the feasibility of using intramyocardial injection or intracoronary infusions of recombinant adeno-a ... | 1999 | 9892583 |
| human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2. | adeno-associated virus 2 (aav)-based vectors have gained attention as a potentially useful alternative to the more commonly used retroviral and adenoviral vectors for human gene therapy. although aav uses the ubiquitously expressed cell surface heparan sulfate proteoglycan (hspg) as a receptor, the transduction efficiency of aav vectors varies greatly in different cells and tissues in vitro and in vivo. we demonstrate here that cell surface expression of hspg alone is insufficient for aav infect ... | 1999 | 9883842 |
| human adeno-associated virus type 5 is only distantly related to other known primate helper-dependent parvoviruses. | we have characterized 95% (4,404 nucleotides) of the genome of adeno-associated virus type 5 (aav5), including part of the terminal repeats and the terminal resolution site. our results show that aav5 is different from all other described aav serotypes at the nucleotide level and at the amino acid level. the sequence homology to aav2, aav3b, aav4, and aav6 at the nucleotide level is only between 54 and 56%. the positive strand contains two large open reading frames (orfs). the left orf encodes t ... | 1999 | 9882294 |
| site-specific targeting of dna plasmids to chromosome 19 using aav cis and trans sequences. | 2000 | 10561835 | |
| adeno-associated virus based gene therapy in skeletal muscle. | 2000 | 10561836 | |
| aspartoacylase gene transfer to the mammalian central nervous system with therapeutic implications for canavan disease. | with the ultimate goal of developing safe and effective in vivo gene therapy for the treatment of canavan disease and other neurological disorders, we developed a non-viral lipid-entrapped, polycation-condensed delivery system (lpd) for central nervous system gene transfer, in conjunction with adeno-associated virus (aav)-based plasmids containing recombinant aspartoacylase (aspa). the gene delivery system was tested in healthy rodents and primates, before proceeding to preliminary studies in 2 ... | 2000 | 10894213 |
| kinetics of recombinant adeno-associated virus-mediated gene transfer. | recombinant adeno-associated virus (raav) vectors have been shown to be useful for efficient gene delivery to a variety of dividing and nondividing cells. mechanisms responsible for the long-term, persistent expression of the raav transgene are not well understood. in this study we investigated the kinetics of raav-mediated human factor ix (hfix) gene transfer into human primary myoblasts and myotubes. transduction of both myoblasts and myotubes occured with a similar and high efficiency. after ... | 2000 | 10729130 |
| transduction of hepatocellular carcinoma (hcc) using recombinant adeno-associated virus (raav): in vitro and in vivo effects of genotoxic agents. | adeno-associated virus (aav) is an attractive tool for gene therapy. here we investigated the in vitro and in vivo transduction of hepatocellular carcinoma (hcc) cells by an aav vector and the efficacy of different strategies to enhance the transduction of the tumor. | 2000 | 10898318 |
| nonrandom transduction of recombinant adeno-associated virus vectors in mouse hepatocytes in vivo: cell cycling does not influence hepatocyte transduction. | recombinant adeno-associated virus vectors (raav) show promise in preclinical trials for the treatment of genetic diseases including hemophilia. liver-directed gene transfer results in a slow rise in transgene expression, reaching steady-state levels over a period of 5 weeks concomitant with the conversion of the single-stranded raav molecules into high-molecular-weight concatemers in about 5% of hepatocytes. immunohistochemistry and rna in situ hybridization show that the transgene product is m ... | 2000 | 10729154 |
| adeno-associated virus type 5 (aav5) but not aav2 binds to the apical surfaces of airway epithelia and facilitates gene transfer. | in the genetic disease cystic fibrosis, recombinant adeno-associated virus type 2 (aav2) is being investigated as a vector to transfer cftr cdna to airway epithelia. however, earlier work has shown that the apical surface of human airway epithelia is resistant to infection by aav2, presumably as a result of a lack of heparan sulfate proteoglycans on the apical surface. this inefficiency can be overcome by increasing the amount of vector or by increasing the incubation time. however, these interv ... | 2000 | 10729159 |
| adeno-associated virus vector carrying human minidystrophin genes effectively ameliorates muscular dystrophy in mdx mouse model. | duchenne muscular dystrophy (dmd) is the most common and lethal genetic muscle disorder, caused by recessive mutations in the dystrophin gene. one of every 3,500 males suffers from dmd, yet no treatment is currently available. genetic therapeutic approaches, using primarily myoblast transplantation and adenovirus-mediated gene transfer, have met with limited success. adeno-associated virus (aav) vectors, although proven superior for muscle gene transfer, are too small (5 kb) to package the 14-kb ... | 2000 | 11095710 |
| principles of gene therapy: potential applications in the treatment of cerebral ischaemia. | in this review we explore gene therapy as a possible treatment for conditions causing cerebral ischaemia and briefly consider other neurological pathologies such as brain tumours. dna transfer may be achieved using retrovirus, herpes simplex virus, adenovirus, and adeno-associated virus vectors or liposomes. after cerebral ischaemia, these vectors are used to upregulate genes that increase survival and inhibit those that promote death in the injured cells. in contrast, in brain tumours gene ther ... | 2000 | 11198761 |
| scaleable chromatographic purification process for recombinant adeno-associated virus (raav). | adeno-associated virus (aav) is a human parvovirus currently being developed as a vector for gene therapy applications. traditionally aav has been purified from cell lysates using cscl gradients; this approach however is not likely to be useful in large-scale manufacturing. moreover gradient-purified aav vectors tend to be contaminated with significant levels of cellular and adenoviral proteins and nucleic acid. to address the issue of purification we have developed a process scale method for th ... | 2000 | 11199265 |
| adeno-associated virus production of soluble tumor necrosis factor receptor neutralizes tumor necrosis factor alpha and reduces arthritis. | the major limitation of adenovirus is its association with induction of an inflammatory response and relatively short-term production of the gene therapy transgene product. adeno-associated virus (aav) is a 4.68-kb single-strand dna virus that contains itrs for viral replication and a packaging signal, and also has been engineered to contain therapeutic genes up to 5 kb in length. transduction of recombinant aav (raav) results in low inflammatory response and long-term expression. we have cloned ... | 2000 | 11096446 |