Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
|---|
| adeno-associated virus vector mediated gene transfer to pancreatic beta cells. | insulin-dependent diabetes mellitus (iddm) or type 1 diabetes is an autoimmune disease that results in destruction of the insulin-producing pancreatic islet beta cells. several factors induce the invasion of immune cells into islets and trigger inflammation. gene therapy approaches targeting the islet cells could be an effective treatment to prevent the onset or reverse type 1 diabetes. allogeneic islet transplantation provides short-term treatment. however, genetically modified islets, which re ... | 2000 | 11021593 |
| adeno-associated virus rnas appear in a temporal order and their splicing is stimulated during coinfection with adenovirus. | we have used a quantitative rnase protection assay to characterize the relative accumulation and abundance of individual adeno-associated virus type 2 (aav) rnas throughout the course of aav-adenovirus coinfections and preinfections. we have demonstrated that there is a previously unrecognized temporal order to the appearance of aav rnas. first, unspliced p5-generated transcripts, which encode rep78, were detectable prior to the significant accumulation of other aav rnas. ultimately, as previous ... | 2000 | 11024114 |
| adeno-associated virus-mediated vascular endothelial growth factor gene transfer into cardiac myocytes. | vascular endothelial growth factor (vegf) is an angiogenic growth factor that stimulates endothelial cell proliferation, increases endothelial permeability, and promotes collateral vessel formation. we transferred human vegf gene into rat cardiac myocytes using adeno-associated virus (aav) vectors and investigated whether vegf secreted from the transduced cardiac myocytes promoted proliferation of endothelial cells. we produced vegf-expressing aav vectors (aav-vegf) by the adenovirus-free method ... | 2000 | 11026643 |
| self-amplification system for recombinant adeno-associated virus production. | a recently reported system for recombinant adeno-associated virus (raav) production does not require infection of a helper virus and depends on the transfection with a huge amount of three plasmids: aav-vector, aav-helper, and adenovirus-helper plasmids. toward simplifying raav production, as a first step, we tested the use of the raav itself instead of the aav-vector plasmid as a source of raav dna and determined the optimal timing of infection and dose of the input raav. when 293 cells were in ... | 2000 | 11027513 |
| the adeno-associated virus vector for orthopaedic gene therapy. | during the last decade researchers working with recombinant adeno-associated virus have shown the use of this vector for efficient and long-term gene transfer in various tissues including lung, muscle, brain, spinal cord, retina, and liver. in 1999 the first results documenting the use of this vector in transducing joint cells were published. additional advantages of recombinant adeno-associated virus for in vivo gene therapy are: (1) its ability to transduce nondividing cells; (2) site-specific ... | 2000 | 11039749 |
| gene therapy approaches for treating rheumatoid arthritis. | current gene therapy approaches for treating rheumatoid arthritis have made use of gene transfer technology as an improved delivery system for emerging proteins and other biologicals whose activities may have therapeutic value. preclinical research has focused on two primary directions, evaluation of methods of gene delivery and identification of gene products with antiarthritic potential. although there are reports involving systemic gene delivery, the bulk of effort has focused on local, intra ... | 2000 | 11039782 |
| adeno-associated virus (aav) rep protein enhances the generation of a recombinant mini-adenovirus (ad) utilizing an ad/aav hybrid virus. | mini-adenoviruses (mad) deleted of all viral coding regions represent an emerging approach for transgene expression. we have exploited the unique features of the adeno-associated virus (aav) terminal repeats within the context of an adenovirus-adeno-associated hybrid virus (ad/aav) as a strategy for rapid and efficient generation of mad. excision and generation of mad from the parental ad/aav hybrid vector was achieved in 293 cells through recombination but without selection for mad production. ... | 2000 | 11044082 |
| site-specific integration of an adeno-associated virus vector plasmid mediated by regulated expression of rep based on cre-loxp recombination. | recombinant adeno-associated virus (aav) type 2 has attracted attention because it appears to have the potential to serve as a vector for human gene therapy. an interesting feature of wild-type aav is its site-specific integration into aavs1, a defined locus on chromosome 19. this reaction requires the presence of two viral elements: inverted terminal repeats and rep78/68. accordingly, current aav vectors lacking the rep gene lack the capacity for site-specific integration. in this report, we de ... | 2000 | 11044107 |
| recombinant adeno-associated virus vector-based gene transfer for defects in oxidative metabolism. | defects in oxidative metabolism may be caused by mutations either in nuclear genes or in mitochondrial dna (mtdna). we tested the hypothesis that recombinant adeno-associated virus (raav) could be used to complement mtdna mutations. aav vector constructs were designed to express the reporter gene encoding green fluorescent protein (gfp), fused to a targeting presequence that directed gfp to be translocated into mitochondria. these vectors mediated expression of mitochondrial-localized gfp, as in ... | 2000 | 11044909 |
| purification of recombinant adeno-associated virus vectors by column chromatography and its performance in vivo. | recombinant adeno-associated virus (aav) holds much promise for human gene therapy. while evidence indicates that aav mediates long-term gene transfer in several different tissues, difficulty in preparing and purifying this viral vector in large quantities remains a major obstacle for evaluating aav vectors in clinical trials. the current method of purification, based on sedimentation through cesium chloride, is not scaleable and yields product of insufficient quality. in this article we report ... | 2000 | 11044910 |
| chronic ethanol increases adeno-associated viral transgene expression in rat liver via oxidant and nfkappab-dependent mechanisms. | recombinant adeno-associated virus (raav) transduction is limited in vivo, yet can be enhanced by hydroxyurea, ultraviolet-irradiation, or adenovirus coinfection, possibly via mechanisms involving stress in the host cell. because chronic ethanol induces oxidative stress, it was hypothesized that chronic ethanol would increase raav transduction in vivo. to test this hypothesis, raav encoding beta-galactosidase was given to wistar rats that later received either ethanol diet or high-fat control di ... | 2000 | 11050056 |
| adeno-associated virus vector-mediated gene transfer to somatic cells in the central nervous system. | 2000 | 11050954 | |
| production of recombinant adeno-associated virus. | currently, raav appears to be one of the most promising vectors for gene therapy applications. attractive features of the vector include nonpathogenicity, the ability to infect nondividing cells, escape from host immune responses, and integration into the host genome. tremendous progress has been made in the production of this vector, which makes it possible to start to examine the vector performance in large animals and to implement the transition to phase i human clinical trials with a variety ... | 2000 | 11050955 |
| current status of viral gene therapy for brain tumours. | malignant glial tumours represent the majority of primary brain tumours. despite the use of many adjunctive treatment strategies in addition to surgery, the prospect of cure or even long-term survival is poor. in the last decade, there has been an explosion of interest in the development of delivery systems that will allow the expression of exogenous genes in the cns. for the most part, these systems are based upon modified viruses. to date, the greatest experience has been with retroviruses, he ... | 2000 | 11060704 |
| critical aspects of viral vectors for gene transfer into the kidney. | viral vectors have been used in vitro and in vivo for more than a decade, with some significant results in specific situations, e.g., when recombinant adeno-associated virus is used for the long-term transduction of skeletal muscle in coagulation factor ix-deficient patients. however, the kidney has been quite difficult to transduce with any viral vector currently available. when viral transduction occurs, it is often heterogeneous, transient, and eventually associated with immune and toxic side ... | 2000 | 11065348 |
| adeno-associated virus vectors for gene therapy: more pros than cons? | gene therapy vectors based on the adeno-associated virus (aav) are being developed for a widening variety of therapeutic applications. enthusiasm for aav is due, not only to the relative safety of these vectors, but also to advances in understanding of the unique biology of this virus. this review examines a number of long-standing concerns regarding the utility of aav for gene transfer in light of many new insights into the biology, immunology and production of aav. | 2000 | 11074369 |
| in vivo transduction of cerebellar purkinje cells using adeno-associated virus vectors. | we investigated whether adenovirus or adeno-associated virus vectors can transduce cerebellar purkinje cells (pcs) in vivo. mice were injected in the deep cerebellar nuclei (dcn) with lacz-transducing adenovirus (ad.rsv-betagal) or a recombinant aav serotype 2 (raav2) vector (vtr-cmvbeta) mixed with wild-type adenovirus type 5 (ad5). one week later, ad.rsv-betagal transduced cells were found throughout the cerebellar white matter in a dose-dependent manner, but few transduced pcs were evident. i ... | 2000 | 11082318 |
| inhibition of recombinant adeno-associated virus (raav) transduction by bronchial secretions from cystic fibrosis patients. | the conducting airways are the primary target for gene transfer in cystic fibrosis (cf), yet the inflammation associated with cf lung disease could potentially pose a significant barrier to gene transfer vectors, such as recombinant adeno-associated virus (raav). in order to investigate this possibility, aliquots of bronchoalveolar lavage (bal) fluid from eight individuals with cf were tested for their in vitro inhibitory effects on raav transduction, along with bal from non-cf individuals. whil ... | 2000 | 11083501 |
| recombinant adeno-associated virus vector expressing glial cell line-derived neurotrophic factor reduces ischemia-induced damage. | to explore the potential of using the recombinant adeno-associated viral (raav) vector, expressing glial cell line-derived neurotrophic factor (gdnf) as the gene therapy for stroke, we injected raav vectors expressing gdnf (raav-gdnf) into the cortex of rats which had been experiencing transient bilateral common carotid artery ligation and right middle cerebral artery ligation for 90 min. gdnf levels in cortical tissues of raav-gdnf-injected animals were significantly higher than in the control ... | 2000 | 11085892 |
| the adenovirus e4 orf6 and e1b 55 kda proteins cooperate in a p53-independent manner to enhance transduction by recombinant adeno-associated virus vectors. | the observation that exposure of target cells to genotoxic stress or adenovirus infection enhances recombinant adeno-associated virus (raav) transduction is an important lead towards defining the raav transduction mechanism, and has significant implications for the exploitation of raav in gene therapy applications. the adenovirus-mediated enhancement of raav transduction has been mapped to the e4 orf6 gene, and expression of e4 orf6 alone has been considered necessary and sufficient to mediate t ... | 2000 | 11086129 |
| glucose-responsive gene delivery in pancreatic islet cells via recombinant adeno-associated viral vectors. | recent progress in genetic engineering presents the possibility of providing physiologically regulated glucose metabolism in individuals with diabetes. the objective of this study is to explore the feasibility of obtaining glucose dependent gene expression in the pancreatic beta-cell lines via recombinant adeno-associated virus type 2 (raav) mediated gene transfer. | 2000 | 11087036 |
| high-titer, wild-type free recombinant adeno-associated virus vector production using intron-containing helper plasmids. | recombinant adeno-associated virus (raav) is capable of directing long-term, high-level transgene expression without destructive cell-mediated immune responses. however, traditional packaging methods for raav vectors are generally inefficient and contaminated with replication-competent aav (rcaav) particles. although wild-type aav is not associated with any known human diseases, contaminating rcaav particles may affect raav gene expression and are an uncontrolled variable in many aav gene transf ... | 2000 | 11090141 |
| efficient replication of adeno-associated virus type 2 vectors: a cis-acting element outside of the terminal repeats and a minimal size. | recombinant adeno-associated virus type 2 (aav2) can be produced in adenovirus-infected cells by cotransfecting a plasmid containing the recombinant aav2 genome, which is generally comprised of the viral terminal repeats flanking a transgene, together with a second plasmid expressing the aav2 rep and cap genes. however, recombinant viruses generally replicate inefficiently, often producing 100-fold fewer virus particles per cell than can be obtained after transfection with a plasmid containing a ... | 2000 | 11090148 |
| 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 |
| 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 |
| long-term real-time monitoring of adeno-associated virus-mediated gene expression in the rat retina. | previous studies have demonstrated that adeno-associated virus (aav) efficiently transduced retinal pigmented epithelial (rpe) cells. the goal of this study was to further evaluate and characterize transgene expression within the rpe cells over time in vivo. | 2000 | 11097287 |
| remission in models of type 1 diabetes by gene therapy using a single-chain insulin analogue. | a cure for diabetes has long been sought using several different approaches, including islet transplantation, regeneration of beta cells and insulin gene therapy. however, permanent remission of type 1 diabetes has not yet been satisfactorily achieved. the development of type 1 diabetes results from the almost total destruction of insulin-producing pancreatic beta cells by autoimmune responses specific to beta cells. standard insulin therapy may not maintain blood glucose concentrations within t ... | 2000 | 11100731 |
| viral vectors for gene delivery and gene therapy within the endocrine system. | the transfer of genetic material into endocrine cells and tissues, both in vitro and in vivo, has been identified as critical for the study of endocrine mechanisms and the future treatment of endocrine disorders. classical methods of gene transfer, such as transfection, are inefficient and limited mainly to delivery into actively proliferating cells in vitro. the development of viral vector gene delivery systems is beginning to circumvent these initial setbacks. several kinds of viruses, includi ... | 2000 | 10657846 |
| a chimeric protein containing the n terminus of the adeno-associated virus rep protein recognizes its target site in an in vivo assay. | the rep78 and rep68 proteins of adeno-associated virus (aav) type 2 are involved in dna replication, regulation of gene expression, and targeting site-specific integration. they bind to a specific rep recognition sequence (rrs) found in both the viral inverted terminal repeats and the aavs1 integration locus on human chromosome 19. previous in vitro studies implied that an n-terminal segment of rep is involved in dna recognition, while additional domains might stabilize binding and mediate multi ... | 2000 | 10666268 |
| humoral immunity to adeno-associated virus type 2 vectors following administration to murine and nonhuman primate muscle. | adeno-associated virus (aav) is being developed as a vector capable of conferring long-term gene expression, which is useful in the treatment of chronic diseases. in most therapeutic applications, it is necessary to readminister the vector. this study characterizes the humoral immune response to aav capsid proteins following intramuscular injection and its impact on vector readministration. studies of mice and rhesus monkeys demonstrated the formation of neutralizing antibodies to aav capsid pro ... | 2000 | 10666273 |
| adeno-associated virus major rep78 protein disrupts binding of tata-binding protein to the p97 promoter of human papillomavirus type 16. | adeno-associated virus type 2 (aav) is known to inhibit the promoter activities of several oncogenes and viral genes, including the human papillomavirus type 16 (hpv-16) e6 and e7 transforming genes. however, the target elements of aav on the long control region (lcr) upstream of e6 and e7 oncogenes are elusive. a chloramphenicol acetyltransferase assay was performed to study the effect of aav on the transcription activity of the hpv-16 lcr in siha (hpv-positive) and c-33a (hpv-negative) cells. ... | 2000 | 10666281 |
| transduction and utility of the granulocyte-macrophage colony-stimulating factor gene into monocytes and dendritic cells by adeno-associated virus. | the genetic manipulation of antigen-presenting dendritic cells (dc) offers promise for stimulating the immune response, in particular for anticancer and antiviral protocols. as adeno-associated virus (aav) has shown promise as a gene delivery vector for transducing a variety of hematopoietic cell types, we have investigated aav's ability to genetically alter dc. in this analysis, we modified the standard granulocyte-macrophage colony-stimulating factor (gm-csf) and interleukin-4 (il-4) treatment ... | 2000 | 10670649 |
| developments in gene therapy for muscular dystrophy. | gene therapy for muscular dystrophy (md) presents significant challenges, including the large amount of muscle tissue in the body, the large size of many genes defective in different muscular dystrophies, and the possibility of a host immune response against the therapeutic gene. overcoming these challenges requires the development and delivery of suitable gene transfer vectors. encouraging progress has been made in modifying adenovirus (ad) vectors to reduce immune response and increase capacit ... | 2000 | 10679969 |
| aav vectors: is clinical success on the horizon? | potential applications and impact of the adeno-associated virus (aav) as a gene transfer vector have expanded rapidly in the last decade. recent advances in the production of high-titer purified raav vector stocks have made the transition to human clinical trials a reality in the last moments of the millenium. production improvements will be complemented in the coming years with understanding of and innovations in the targeting and packaging of raav, the design of transgene cassettes, and the ho ... | 2000 | 10680012 |
| a role for single-stranded templates in cell-free adeno-associated virus dna replication. | assays have been described in which duplex adeno-associated virus (aav) dna can be replicated in hela cell extracts with exogenous aav rep protein. these assays appear to mimic the aav dna replication that occurs in the cell, including the ability of extracts from adenovirus (ad)-infected cells to replicate duplex aav dna templates more efficiently than extracts from uninfected cells can. we showed previously that the ad-infected extract was able to support a more processive replication than the ... | 2000 | 10623736 |
| impaired intracellular trafficking of adeno-associated virus type 2 vectors limits efficient transduction of murine fibroblasts. | although adeno-associated virus type 2 (aav) has gained attention as a potentially useful alternative to the more commonly used retrovirus- and adenovirus-based vectors for human gene therapy, efficient gene transfer and transgene expression by aav vectors require that the following two obstacles be overcome. first, the target cell must express the receptor and the coreceptor for aav infection, and second, the cell must allow for viral second-strand dna synthesis. we now describe a third obstacl ... | 2000 | 10623762 |
| full functional rescue of a complete muscle (ta) in dystrophic hamsters by adeno-associated virus vector-directed gene therapy. | limb girdle muscular dystrophy (lgmd) 2f is caused by mutations in the delta-sarcoglycan (sg) gene. previously, we have shown successful application of a recombinant adeno-associated virus (aav) vector for genetic and biochemical rescue in the bio14.6 hamster, a homologous animal model for lgmd 2f (j. li et al., gene ther. 6:74-82, 1999). in this report, we show efficient and long-term delta-sg expression accompanied by nearly complete recovery of physiological function deficits after a single-d ... | 2000 | 10627554 |
| repeat transduction in the mouse lung by using adeno-associated virus vectors with different serotypes. | vectors derived from adeno-associated virus type 2 (aav2) promote gene transfer and expression in the lung; however, we have found that while gene expression can persist for at least 8 months in mice, it was reduced dramatically in rabbits over a period of 2 months. the efficiency and persistence of aav2-mediated gene expression in the human lung have yet to be determined, but it seems likely that readministration will be necessary over the lifetime of an individual. unfortunately, we have found ... | 2000 | 10627564 |
| adeno-associated virus type 2 nonstructural protein rep78 suppresses translation in vitro. | adeno-associated virus type 2 nonstructural protein rep78 [621 amino acids (aa) long] affects the expression of various cellular and viral genes. in this study we examined the effects of rep78 on expression of the luciferase gene from the human cytomegalovirus immediate-early promoter in hela cells and on translation of rna encoding luciferase in rabbit reticulocyte lysate. when rep78 and luciferase were coexpressed, the luciferase activity decreased despite increased levels of luciferase mrna i ... | 2000 | 10612674 |
| adenoviral and adeno-associated viral transfer of genes to the peripheral nervous system. | targeted expression of foreign genes to the peripheral nervous system is interesting for many applications, including gene therapy of neuromuscular diseases, neuroanatomical studies, and elucidation of mechanisms of axonal flow. here we describe a microneurosurgical technique for injection of replication-defective viral vectors into dorsal root ganglia (drg). adenovirus- and adeno-associated virus-based vectors with transcriptional competence for drg neurons led to expression of the gene of inte ... | 2000 | 10618437 |
| conditional site-specific integration into human chromosome 19 by using a ligand-dependent chimeric adeno-associated virus/rep protein. | it is of great interest for gene therapy to develop vectors that drive the insertion of a therapeutic gene into a chosen specific site on the cellular genome. adeno-associated virus (aav) is unique among mammalian viruses in that it integrates into a distinct region of human chromosome 19 (integration site aavs1). the inverted terminal repeats (itrs) flanking the aav genome and the aav-encoded nonstructural proteins rep78 and/or rep68 are the only viral elements necessary and sufficient for site ... | 2000 | 10590116 |
| incorporation of adeno-associated virus in a calcium phosphate coprecipitate improves gene transfer to airway epithelia in vitro and in vivo. | adeno-associated virus (aav) is inefficient at infecting differentiated airway epithelia because of a lack of receptors at the apical surface. we hypothesized that incorporation of aav in a calcium phosphate coprecipitate would circumvent this barrier. interestingly, coprecipitation of aav type 2 improved gene transfer to differentiated human airway epithelia in vitro and to the mouse lung in vivo. these results suggest that delivery of aav as a cap(i) coprecipitate may significantly enhance its ... | 2000 | 10590145 |
| epitope mapping of human anti-adeno-associated virus type 2 neutralizing antibodies: implications for gene therapy and virus structure. | recombinant adeno-associated virus type 2 (aav) is a common vector used in human gene therapy protocols. we characterized the humoral immune response to aav and observed that 80% of normal human subjects have anti-aav antibodies and that 18% have neutralizing antibodies. to analyze the effect of neutralizing antibodies on aav readministration, we attempted to deliver recombinant aav expressing human factor ix (aav-hfix) intraportally into the livers of mice which had been preexposed to aav and s ... | 2000 | 10644347 |
| antivector and antitransgene host responses in gene therapy. | current viral gene therapy vectors effectively transfer genes in vivo at the price of eliciting innate and acquired host responses against the vector and/or transgene. antigens present in the viral vector and the expression of the transgene both cause cellular and humoral immune responses dependent on the viral vector, the route of administration, and the genotype and infection history of the host. in general, adenoviral vectors cause strong immune responses, which result in only transient expre ... | 2000 | 11249767 |
| micro-injection-mediated hematopoietic stem cell gene therapy. | over the past decade, significant attention has been devoted to the development of viral vectors (i.e., retrovirus, lentivirus, adeno-associated virus) and conditions capable of transducing hematopoietic stem cells. after several years of disappointing results, recent reports in humans and other primates, most particularly the french report of successful treatment of x-linked severe combined immune deficiency (scid) [1.], indicate that viral approaches will be successful in treating specific hem ... | 2000 | 11249771 |
| herpes simplex virus-mediated gene transfer as a tool for neuropsychiatric research. | there is an enormous initiative to establish causal relationships between brain biology (including patterns of gene expression) and behavior. unfortunately, genetic intervention is not accomplished easily in the brain. one strategy is to engineer and deliver to the brain specialized viral vectors that carry a gene (or genes) of interest, thereby exploiting the natural ability of viruses to insert genetic information into cells. when delivered to the brain, these vectors cause infected cells to i ... | 2000 | 11253955 |
| [construction of a hepatoma-targeting vector of adeno-associated virus containing human alpha-fetoprotein promoter and wild p53 gene in gene therapy of liver cancer]. | to construct plasmids that express target genes in hepatoma cell line using adeno-associated virus (aav) vectors containing human afp promoter. | 2000 | 11798803 |
| 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 |
| size does matter: overcoming the adeno-associated virus packaging limit. | recombinant adeno-associated virus (raav) vectors mediate long-term gene transfer without any known toxicity. the primary limitation of raav has been the small size of the virion (20 nm), which only permits the packaging of 4.7 kilobases (kb) of exogenous dna, including the promoter, the polyadenylation signal and any other enhancer elements that might be desired. two recent reports (d duan et al: nat med 2000, 6:595-598; z yan et al: proc natl acad sci usa 2000, 97:6716-6721) have exploited a u ... | 2000 | 11667959 |
| 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 | |
| [effect of 6a8 alpha-mannosidase expression on the proliferative response of human b cell line 3d5]. | to study the effect of 6a8 alpha-mannosidase expression on the proliferative response of human b cell 3d5. | 2000 | 12903396 |
| developing protocols for recombinant adeno-associated virus-mediated gene therapy in space. | with the advent of the era of international space station (iss) and mars exploration, it is important more than ever to develop means to cure genetic and acquired diseases, which include cancer and aids, for these diseases hamper human activities. thus, our ultimate goal is to develop protocols for gene therapy, which are suitable to humans on the earth as well as in space. specifically, we are trying to cure the hemoglobinopathies, beta-thalassemia (cooley's anemia) and sickle cell anemia, by g ... | 2000 | 12697549 |
| scaling-up production of recombinant aav vectors for clinical applications. | recombinant adeno-associated virus (aav)-based vectors capable of expressing therapeutic gene products in vivo have shown significant promise for human gene therapy. a major challenge for these applications is the development of processes to enable production of large quantities of aav vectors and purification of material that is well characterized and appropriate for parenteral administration. several cell culture systems have been developed for aav vector production, and a limited number of th ... | 2000 | 19649903 |
| group i avian adenovirus and avian adeno-associated virus in turkey poults with inclusion body hepatitis. | adenoviral inclusion body hepatitis (ibh) is rare in turkeys. avian adenovirus group i and avian adeno-associated virus were isolated from the liver and pooled intestinal samples from 4-week-old turkey poults on two different ranches experiencing increased mortality. grossly, a few birds from each ranch had a slightly enlarged liver with white foci of necrosis randomly scattered throughout. microscopically, there was coagulative necrosis of hepatocytes with infiltration of a mixed population of ... | 2001 | 19184960 |
| adeno-associated virus-mediated transfer of endothelial nitric oxide synthase gene reduces the vasoconstrictive response. | adeno-associated virus (aav) has a number of attractive features for gene therapy including the ability to transduce nondividing cells and long term transgene expression. | 2001 | 20428445 |
| [the study of transfected cardiomyocytes by recombinant adenovirus and adeno-associated virus]. | recombinant adenovirus (rad) and adeno-associated virus (raav) were created, in which beta2-adrenergic receptors (beta2-ar) gene is under control of the cmv promotor, the cultured neonate rat ventricular myocytes were infected by the two vectors, and the expression of beta2-ar on cultured neonate rat ventricular myocytes was assessed. | 2001 | 21171405 |
| generation of recombinant adeno-associated virus. | adeno-associated virus (aav) was discovered about 30 yr ago as a contaminant of adenovirus preparations. since its discovery, researchers have described many unique characteristics of aav biology that have made it attractive as a potential vector for gene therapy. for example, aav is not pathogenic, approx 80% of adults in the united states are seropositive, but in no case has the virus been implicated as the etiological agent for a human disease. aav is a defective parvovirus with a single-stra ... | 2001 | 21394584 |
| [growth inhibition of human nasopharyngeal carcinoma cell cne-2l2 caused by suppression of 6a8 alpha-mannosidase expression]. | to study the relationship between 6a8 alpha-mannosidase expression and growth of cne-2l2 cells, a human nasopharyngeal carcinoma cell line. | 2001 | 12905853 |
| dna of adeno-associated virus (aav) in testicular tissue and in abnormal semen samples. | human genital tissues, including spermatozoa, have been found to be frequently infected with the helper-virus dependent parvovirus, adeno-associated virus (aav). | 2001 | 11679515 |
| [possibility and future problems of gene therapy for gastric cancer]. | recently, stage-oriented surgery has been performed for gastric cancer, but a new strategy is necessary for stage iv gastric cancer. the first target of gene therapy for gastric cancer was for stage iv patients with-widespread lymph node metastases and/or peritoneal dissemination. we reported on suicide gene therapy in experimental gastric cancer induced by enng in the dog, and the results showed that in situ gene transfer of a suicide gene (ad. caghsv-tk) followed by prodrug (gcv) treatment may ... | 2001 | 11681005 |
| expression of aav rep proteins in sv40-transformed and untransformed cells: reciprocal interaction with host dna synthesis. | adeno-associated virus (aav) inhibits the induction of host dna synthesis by simian virus 40 (sv40) large-tumour (t) antigen, mediated through aav-encoded 'rep' regulatory proteins. rep proteins are normally synthesized by aav-infected cells only in the presence of adenovirus. however, we observed a low level of rep protein expression in sv40 transformed cells even in the absence of helper virus. in an effort to understand the functional interaction between sv40 t antigen and regulators of aav r ... | 2001 | 11684891 |
| recombinant adeno-associated virus vectors for cystic fibrosis gene therapy. | cystic fibrosis (cf) is an autosomal recessive inherited disorder that affects approximately 30,000 north americans. defects in the cf transmembrane conductance regulator (cftr) gene lead to altered secretions from exocrine glands and the pulmonary airways, to a heightened susceptibility to airway infections with pseudomonas aeruginosa, and to severe airway inflammation. early attempts to develop a genetic therapy for cf have not met with great clinical success, but these efforts have driven the ... | 2001 | 11699895 |
| technology evaluation: anticancer gene therapy, medigene. | medigene is developing recombinant adeno-associated virus (raav) vectors for the potential treatment of cancer, and in particular, melanoma and ovarian cancer [304021]. in june 2001, medigene and its collaborator aventis pharma announced the start of a phase i/ii trial for a vaccine against malignant melanoma [413513]. the therapy works by boosting the patient's immune response. autologous or allogeneic tumor cells are transduced with recombinant adeno-associated virus (raav) vectors, which expr ... | 2001 | 11699897 |
| molecular regulation and biological function of adenovirus early genes: the e4 orfs. | over the past few years there have been a number of interesting advances in our understanding of the functions encoded by the adenovirus early transcription unit 4 (ad e4). a large body of recent data demonstrates that e4 proteins encompass an unexpectedly diverse collection of functions required for efficient viral replication. e4 gene products operate through a complex network of protein interactions with key viral and cellular regulatory components involved in transcription, apoptosis, cell c ... | 2001 | 11707318 |
| adeno-associated virus (aav) site-specific recombination does not require a rep-dependent origin of replication within the aav terminal repeat. | adeno-associated virus (aav) is the only known eukaryotic virus capable of targeted integration in human cells. an aav rep binding element (rbe) and terminal resolution site (trs) identical to the viral terminal repeats required for aav dna replication are located on chromosome (ch) 19. both ch-19 rbe and trs elements have been shown to be essential for viral targeting to this locus. to characterize the role of the aav inverted terminal repeat (itr) cis-acting sequences in targeted integration a ... | 2001 | 11707592 |
| long-term protection of retinal structure but not function using raav.cntf in animal models of retinitis pigmentosa. | the present study aimed to determine whether intravitreal administration of an adeno-associated virus (aav) carrying ciliary neurotrophic factor (cntf) can achieve long-term morphological and physiological rescue of photoreceptors in animal models of retinitis pigmentosa, and whether injection of this virus after degeneration begins is effective in protecting the remaining photoreceptors. we injected raav.cntf.gfp intravitreally in early postnatal prph2(rd2/rd2) (formerly rds/rds) mice and in ad ... | 2001 | 11708883 |
| involvement of cellular double-stranded dna break binding proteins in processing of the recombinant adeno-associated virus genome. | unlike postmitotic tissues in vivo, transduction of cultured cells is poor with recombinant adeno-associated virus (raav). the ability of raav to transduce cells is greatly enhanced by a variety of agents that induce dna damage and is elevated in cells defective in the ataxia telangiectasia gene product (atm), showing increased genomic instability. here we show that dna double-stranded break (dsb) repair pathways are involved in the regulation of raav transduction efficiency. by quantitative chr ... | 2001 | 11711618 |
| widespread gene delivery and structure-specific patterns of expression in the brain after intraventricular injections of neonatal mice with an adeno-associated virus vector. | developing a system for widespread somatic gene transfer in the central nervous system (cns) would be beneficial for understanding the global influence of exogenous genes on animal models. we injected an adeno-associated virus serotype 2 (aav2) vector into the cerebral lateral ventricles at birth and mapped its distribution and transduction pattern from a promoter capable of expression in multiple targets. the injections resulted in structure-specific patterns of expression that were maintained ... | 2001 | 11711628 |
| electron cryo-microscopy and image reconstruction of adeno-associated virus type 2 empty capsids. | adeno-associated virus type 2 empty capsids are composed of three proteins, vp1, vp2 and vp3, which have relative molecular masses of 87, 72 and 62 kda, respectively, and differ in their n-terminal amino acid sequences. they have a likely molar ratio of 1:1:8 and occupy symmetrical equivalent positions in an icosahedrally arranged protein shell. we have investigated empty capsids of adeno-associated virus type 2 by electron cryo-microscopy and icosahedral image reconstruction. the three-dimensio ... | 2001 | 11713191 |
| evolutionary relationships among parvoviruses: virus-host coevolution among autonomous primate parvoviruses and links between adeno-associated and avian parvoviruses. | the current classification of parvoviruses is based on virus host range and helper virus dependence, while little data on evolutionary relationships among viruses are available. we identified and analyzed 472 sequences of parvoviruses, among which there were (virtually) full-length genomes of all 41 viruses currently recognized as individual species within the family parvoviridae. our phylogenetic analysis of full-length genomes as well as open reading frames distinguished three evolutionary gro ... | 2001 | 11222696 |
| attenuation of hypertension and heart hypertrophy by adeno-associated virus delivering angiotensinogen antisense. | angiotensinogen (agt), one of the major components in the renin-angiotensin system, has been linked to hypertension in humans and animals. we have previously systemically administered antisense oligonucleotides and plasmid vectors with dna that targeted agt and attenuated hypertension in spontaneously hypertensive rats. the aim of the present study was to prolong the effect of antisense treatment by the use of a recombinant adeno-associated viral (raav) vector targeted to agt. using a model of l ... | 2001 | 11230303 |
| aav-mediated delivery of ciliary neurotrophic factor prolongs photoreceptor survival in the rhodopsin knockout mouse. | retinitis pigmentosa (rp), an inherited retinal degenerative disease causing blindness, is characterized by progressive apoptotic death of photoreceptors. therapeutic modification of photoreceptor apoptosis may provide an effective therapy for this disorder. ciliary neurotrophic factor (cntf) has been shown to promote survival of a number of different neuronal cell types, including photoreceptors. the present study aimed to test whether adeno-associated virus (aav)-mediated delivery of the gene ... | 2001 | 11237681 |
| amino-terminal domain exchange redirects origin-specific interactions of adeno-associated virus rep78 in vitro. | the unique ability of adeno-associated virus type 2 (aav) to site-specifically integrate its genome into a defined sequence on human chromosome 19 (aavs1) makes it of particular interest for use in targeted gene delivery. the objective underlying this study is to provide evidence for the feasibility of retargeting site-specific integration into selected loci within the human genome. current models postulate that aav dna integration is initiated through the interactions of the products of a singl ... | 2001 | 11238849 |
| epitope-tagged recombinant aav vectors for expressing neurturin and its receptor in retinal cells. | neurturin (ntn) is a potent neuronal survival factor in the central and peripheral nervous systems. we previously described altered expression of mrnas for ntn and one of its receptor components, gfra-2 in degenerative retinas of rd/rd mice. towards assessing the potential for transfer of these genes to counteract retinal degeneration, we examined recombinant adeno-associated virus (raav) constructs for expression of ntn and gfra-2 transgenes in retinal cells in vitro and for the effect of trans ... | 2001 | 11239244 |
| gene therapy: a 2001 perspective. | in the past year, three clinical trials of gene therapy for haemophilia have been initiated. years of preclinical studies have culminated in translation of research findings into the clinical arena. it is too early to predict which, if any, of these strategies will show efficacy. this paper will review basic aspects of gene therapy for haemophilia and will briefly outline current clinical trials. the three clinical trials all share a dose escalation design. the ongoing trial for haemophilia b in ... | 2001 | 11240615 |
| efficient ex vivo transduction of pancreatic islet cells with recombinant adeno-associated virus vectors. | the ability to transfer immunoregulatory, cytoprotective, or antiapoptotic genes into pancreatic islet cells may allow enhanced posttransplantation survival of islet allografts and inhibition of recurrent autoimmune destruction of these cells in type 1 diabetes. however, transient transgene expression and the tendency to induce host inflammatory responses have limited previous gene delivery studies using viral transfer vectors. we demonstrate here that recombinant adeno-associated virus (raav) s ... | 2001 | 11246870 |
| gamma-rays enhance raav-mediated transgene expression and cytocidal effect of aav-hsvtk/ganciclovir on cancer cells. | adeno-associated virus (aav) vector has several unique properties suited for gene therapy applications. however, relatively low efficiency of transgene expression, which is mainly due to a limited second-strand synthesis from the single-stranded aav genome, can be a problem in some applications that require potent gene expression such as antitumor applications. recently, gamma-ray irradiation has been reported to enhance the second-strand synthesis of the aav genome, and consequently transgene e ... | 2001 | 11263531 |
| adeno-associated virus is associated with a lower risk of high-grade cervical neoplasia. | adeno-associated virus (aav) is a ubiquitous human helper-dependent parvovirus which may interact with human papillomaviruses (hpv) to modify a woman's risk of cervical neoplasia. this analysis was nested in a cohort study of low-income women receiving pap smears as part of their family planning services. we selected cases (55 with high-grade cervical squamous intraepithelial lesions (hsil) and 162 with low-grade lsil) and controls (96 women with normal cervical cytology) and analyzed cervical d ... | 2001 | 11263951 |
| cochlear gene delivery through an intact round window membrane in mouse. | cochlear gene transfer studies in animal models have utilized mainly two delivery methods: direct injection through the round window membrane (rwm) or intracochlear infusion through a cochleostomy. however, the surgical trauma, inflammation, and hearing loss associated with these methods lead us to investigate a less invasive delivery method. herein, we studied the feasibility of a vector transgene-soaked gelatin sponge, gelfoam, for transgene delivery into the mouse cochlea through an intact rw ... | 2001 | 11268286 |
| optimised helper virus-free production of high-quality adeno-associated virus vectors. | clinical development of adeno-associated virus (aav) requires standardised, safe, efficient and scalable procedures for the manufacture of the raav vector, including production, purification and testing. several strategies have been reported for the approach to the manufacturing problem. we report a helper virus-free process that produces high quality raav stocks. | 2001 | 11269337 |
| intracranial injection of recombinant adeno-associated virus improves cognitive function in a murine model of mucopolysaccharidosis type vii. | mucopolysaccharidosis type vii (mps vii) is a lysosomal storage disease caused by the lack of beta-glucuronidase (gusb) activity. gusb deficiency leads to the progressive accumulation of undegraded glycosaminoglycans (gags) in cells of most tissues, including the brain, and is associated with mental retardation. reduction of lysosomal storage in the central nervous system and prevention of cognitive dysfunction may require intracranial delivery of a therapeutic agent during the newborn period th ... | 2001 | 11273777 |
| effect of dna-dependent protein kinase on the molecular fate of the raav2 genome in skeletal muscle. | we report here that the dna-dependent protein kinase (dna-pk) affects the molecular fate of the recombinant adeno-associated virus (raav) genome in skeletal muscle. raav-human alpha1-antitrypsin (raav-haat) vectors were delivered by intramuscular injection to either c57bl/6 (dna-pkcs(+)) or c57bl/6-scid [severe combined immunodeficient (scid), dna-pkcs(-)] mice. in both strains, high levels of transgene expression were sustained for up to 1 year after a single injection. southern blot analysis s ... | 2001 | 11274433 |
| regulated secretion of proinsulin/insulin from human hepatoma cells transduced by recombinant adeno-associated virus. | to employ hepatocytes as surrogate beta-cells for gene therapy of diabetes, a regulatory system was devised in this study by placing the human insulin cdna under the control of the phosphoenolpyruvate carboxykinase (pepck) promoter, followed by the cytomegalovirus immediate early promoter-driven enhanced-green-fluorescent-protein open reading frame. the expression cassette was inserted into the adeno-associated virus vector between two inverted terminal repeats, and used to produce recombinant a ... | 2001 | 11277867 |
| suicide gene therapy for human oral squamous cell carcinoma cell lines with adeno-associated virus vector. | the purpose of this study was to test the possibility of gene transfer as a new therapy for oral cancer. adeno-associated virus (aav) has already been used in the fields of cystic fibrosis and parkinson's disease as a potential vector for gene therapy because of its wide host range, high transduction efficiency, and lack of cytopathogenicity. four human oral squamous cell carcinoma cell lines were transduced with an aav vector containing the beta-galactosidase gene (aavlacz) in vitro. gene trans ... | 2001 | 11287273 |
| adeno-associated virus type 2-mediated gene transfer: altered endocytic processing enhances transduction efficiency in murine fibroblasts. | adeno-associated virus type 2 (aav) is a single-stranded-dna-containing, nonpathogenic human parvovirus that is currently in use as a vector for human gene therapy. however, the transduction efficiency of aav vectors in different cell and tissue types varies widely. in addition to the lack of expression of the viral receptor and coreceptors and the rate-limiting viral second-strand dna synthesis, which have been identified as obstacles to aav-mediated transduction, we have recently demonstrated ... | 2001 | 11287557 |
| recombinant adeno-associated virus (raav) vector-mediated cotransduction of cd70 and cd80 into human malignant melanoma cells results in an additive t-cell response. | genetic modification of malignant melanoma cells by transduction of cdna encoding costimulatory molecules, cytokines or tumor-associated antigens has been shown to induce antitumor immunity. an important step in this scenario is the activation of t cells. cd80 is a pivotal costimulatory molecule for t-cell activation. another molecule with costimulatory activity is cd70. the purpose of this study was to evaluate the capacity of a combined expression of cd70 and cd80 on melanoma cells to amplify ... | 2001 | 11289576 |
| dual level inhibition of e2f-1 activity by adeno-associated virus rep78. | e2f-1, a major cellular transcription factor, plays a pivotal role in regulating the cell cycle. the activity of e2f-1 is negatively regulated by its interaction with retinoblastoma protein (prb), and disruption of the prb-e2f-1 complex, a hallmark of cellular transformation by dna tumor viruses, leads to cell proliferation. adeno-associated virus-2 (aav) is known to have onco-suppressive properties against dna tumor viruses. here we provide, for the first time, the molecular basis for antioncog ... | 2001 | 11294829 |
| integration of adeno-associated virus 2 dna in human mkr melanoma cells induces a peptide with oncosuppressive properties. | integration of adeno-associated virus type 2 (aav2) dna into the genome of the human mkr melanoma cell line grossly alters the cellular phenotype of these cells. cultures derived from aav dna-harboring single cells share various similarities with normal cells in culture, and a considerable number of clones show signs of cellular senescence and terminal differentiation. medium conditioned by such terminally differentiating cells contains a small cytokine-like factor (aav-induced factor [aif]). th ... | 2001 | 11304688 |
| enhanced expression and stable transmission of transgenes flanked by inverted terminal repeats from adeno-associated virus in zebrafish. | mosaic expression of transgenes in the f0 generation severely hinders the study of transient expression in transgenic fish. to avoid mosaicism, enhanced green fluorescent protein (egfp) gene cassettes were constructed and introduced into one-celled zebrafish embryos. these egfp gene cassettes were flanked by inverted terminal repeats (itrs) from adeno-associated virus (aav) and driven by zebrafish alpha-actin (palpha-actin-egfp-itr) or medaka beta-actin promoters (pbeta-actin-egfp-itr). egfp was ... | 2001 | 11307166 |
| two animal models of retinal degeneration are rescued by recombinant adeno-associated virus-mediated production of fgf-5 and fgf-18. | the goal of these experiments was to evaluate the potential of the fibroblast growth factor family members fgf-5 and fgf-18 to rescue photoreceptors from cell death in retinal degenerative disease. two strains of transgenic rats, expressing either a p23h or an s334ter rhodopsin mutation, were used as model systems. the neurotrophic growth factors were delivered by subretinal injection of adeno-associated virus vectors, driving expression of the genes with a constitutive cmv promoter. morphologic ... | 2001 | 11319911 |
| introduction of single base substitutions at homologous chromosomal sequences by adeno-associated virus vectors. | adeno-associated virus (aav) vectors can modify homologous chromosomal sequences at high rates. this gene targeting transduction pathway is distinct from the integrating and episomal pathways used in gene addition approaches. in previous studies, aav vectors were used to introduce small insertion and deletion mutations at homologous chromosomal loci. here we show that aav-mediated gene targeting can also be used to introduce all possible types of single base substitution mutations at the endogen ... | 2001 | 11319913 |
| the role of receptors in the maturation-dependent adenoviral transduction of myofibers. | one of the major hurdles facing the application of adenoviral gene transfer to skeletal muscle is the maturation-dependent transduction of muscle myofibers. it was recently proposed that the viral receptors (coxsackie and adenovirus receptor (car) and the integrins alphavbeta3/beta5) play a major role in the poor adenoviral transduction of mature myofibers. here we report the findings of morphological studies designed to determine experimentally the role of receptors in the adenoviral transducti ... | 2001 | 11320409 |
| gene therapy restores vision in a canine model of childhood blindness. | the relationship between the neurosensory photoreceptors and the adjacent retinal pigment epithelium (rpe) controls not only normal retinal function, but also the pathogenesis of hereditary retinal degenerations. the molecular bases for both primary photoreceptor and rpe diseases that cause blindness have been identified. gene therapy has been used successfully to slow degeneration in rodent models of primary photoreceptor diseases, but efficacy of gene therapy directed at photoreceptors and rpe ... | 2001 | 11326284 |
| adeno-associated virus vector-mediated il-10 gene delivery prevents type 1 diabetes in nod mice. | the development of spontaneous autoimmune diabetes in nonobese diabetic (nod) mice provides for their use as a model of human type 1 diabetes. to test the feasibility of muscle-directed gene therapy to prevent type 1 diabetes, we developed recombinant adeno-associated virus (raav) vectors containing murine cdnas for immunomodulatory cytokines il-4 or il-10. skeletal muscle transduction of female nod mice with il-10, but not il-4, completely abrogated diabetes. raav-il-10 transduction attenuated ... | 2001 | 11717448 |
| transduction of ovarian cancer cells: a recombinant adeno-associated viral vector compared to an adenoviral vector. | recombinant adeno-associated virus (raav) vectors have emerged as vehicles for gene therapy. in addition, anti-neoplastic properties have been attributed to wild-type aav. to take advantage of both features and to overcome technical problems associated with raav preparation, we developed a production method in which raav particles are amplified in an infectious cycle in the presence of wtaav. this results in a 10(3)-10(4)-fold amplification of raav input particles. raav-gfp particles generated b ... | 2001 | 11720450 |
| hot topics in adeno-associated virus as a gene transfer vector. | adeno-associated virus (aav) is a promising viral vector in treating many kinds of hereditary diseases. the broad host range, low level of immune response, and longevity of gene expression observed with this vector have enabled the initiation of a number of clinical trials using this gene delivery system. another potential benefit of aav vectors is their ability to integrate site-specifically in the presence of rep proteins. however, this virus is not well characterized. to obtain high level, pe ... | 2001 | 11721619 |
| gene therapy for hypertension: sense and antisense strategies. | gene therapy for hypertension is needed for the next generation of antihypertensive drugs. current drugs, although effective, have poor compliance, are expensive and short-lasting (hours or one day). gene therapy offers a way to produce long-lasting antihypertensive effects (weeks, months or years). we are currently using two strategies: antisense oligodeoxynucleotides (as-odn), an dantisense dna delivered in viral vectors, to inhibit genes associated with vasoconstrictive properties. it is not ... | 2001 | 11727501 |
| delivery of novel macromolecular drugs against hiv-1. | the development of new low molecular weight drugs against human immunodeficiency virus type 1 (hiv-1) targets other than reverse transcriptase (rt) and protease, such as the integrase and the envelope glycoprotein, is likely to take many years. macromolecular drugs, including antisense oligonucleotides, ribozymes, rna decoys and transdominant mutant proteins, may be able to interfere with a relatively large number of viral targets, thereby decreasing the likelihood of the emergence of drug-resis ... | 2001 | 11728227 |