Publications
| Title | Abstract | Year Filter | PMID(sorted descending) Filter |
|---|
| targeting recombinant adeno-associated virus vectors to enhance gene transfer to pancreatic islets and liver. | human pancreatic islet cells and hepatocytes represent the two most likely target cells for genetic therapy of type i diabetes. however, limits to the efficiency of raav serotype 2 (raav2)-mediated gene transfer have been reported for both of these cell targets. here we report that nonserotype 2 aav capsids can mediate more efficient transduction of islet cells, with aav1 being the most efficient serotype in murine islets, suggesting that receptor abundance could be limiting. in order to test th ... | 2003 | 12907946 |
| [effect of antisense thrombin receptor and p21 double gene co-expression system on the proliferation and apoptosis in human aortic smooth muscle cells]. | to focus on the study of the effect on proliferation and apoptosis of human aortic smooth muscle cells (asmc) by adeno-associated virus (aav) vector carrying antisense thrombin receptor (atr) and p21 double gene co-expression system. | 2002 | 12905648 |
| [inhibition of er alpha-mannosidase expression causes reduction and shortening of microvilli on rat liver epithelial cell wb-f344]. | to study the effect of n-glycosylation on the modification of microvilli on the surface of rat liver epithelial cell wb-f344 and the growth of the cells in culture. | 2003 | 12905608 |
| gene therapy for new bone formation using adeno-associated viral bone morphogenetic protein-2 vectors. | previous reports have suggested that bone morphogenetic protein (bmp) gene therapy could be applied for in vivo bone regeneration. however, these studies were conducted either using immunodeficient animals because of immunogenicity of adenovirus vectors, or using ex vivo gene transfer technique, which is much more difficult to handle. adeno-associated virus (aav) is a replication-defective virus without any association with immunogenicity and human disease. this study was conducted to investigat ... | 2003 | 12883531 |
| modeling cns neurodegeneration by overexpression of disease-causing proteins using viral vectors. | defective handling of proteins is a central feature of major neurodegenerative diseases. the discovery that neuronal dysfunction or degeneration can be caused by mutations in single cellular proteins has given new opportunities to model the underlying disease processes by genetic modification of cells in vitro or by generation of transgenic animals carrying the disease-causing gene. recent developments in recombinant viral-vector technology have opened up an interesting alternative possibility, ... | 2003 | 12850435 |
| receptor targeting of adeno-associated virus vectors. | adeno-associated virus (aav) is a promising vector for human somatic gene therapy. however, its broad host range is a disadvantage for in vivo gene therapy, because it does not allow the selective tissue- or organ-restricted transduction required to enhance the safety and efficiency of the gene transfer. therefore, increasing efforts are being made to target aav-2-based vectors to specific receptors. the studies summarized in this review show that it is possible to target aav-2 to a specific cel ... | 2003 | 12833123 |
| hiv-1 p55gag encoded in the lysosome-associated membrane protein-1 as a dna plasmid vaccine chimera is highly expressed, traffics to the major histocompatibility class ii compartment, and elicits enhanced immune responses. | several genetic vaccines encoding antigen chimeras containing the lysosome-associated membrane protein (lamp) translocon, transmembrane, and cytoplasmic domain sequences have elicited strong mouse antigen-specific immune responses. the increased immune response is attributed to trafficking of the antigen chimera to the major histocompatibility class ii (mhc ii) compartment where lamp is colocalized with mhc ii. in this report, we describe a new form of an hiv-1 p55gag dna vaccine, with the gag s ... | 2003 | 12824194 |
| a biochemical characterization of the adeno-associated virus rep40 helicase. | the human adeno-associated virus (aav) has generated much enthusiasm as a transfer vector for human gene therapy. although clinical gene therapy trials have been initiated using aav vectors, much remains to be learned regarding the basic mechanisms of virus replication, gene expression, and virion assembly. aav encodes four nonstructural, or replication (rep), proteins. the rep78 and rep68 proteins regulate viral dna replication, chromosomal integration, and gene expression. the rep52 and rep40 ... | 2003 | 12824181 |
| preclinical in vivo evaluation of pseudotyped adeno-associated virus vectors for liver gene therapy. | we report the generation and use of pseudotyped adeno-associated viral (aav) vectors for the liver-specific expression of human blood coagulation factor ix (hfix). therefore, an aav-2 genome encoding the hfix gene was cross-packaged into capsids of aav types 1 to 6 using efficient, large-scale technology for particle production and purification. in immunocompetent mice, the resultant vector particles expressed high hfix levels ranging from 36% (aav-4) to more than 2000% of normal (aav-1, -2, and ... | 2003 | 12791653 |
| helper virus-free, optically controllable, and two-plasmid-based production of adeno-associated virus vectors of serotypes 1 to 6. | we present a simple and safe strategy for producing high-titer adeno-associated virus (aav) vectors derived from six different aav serotypes (aav-1 to aav-6). the method, referred to as "hot," is helper virus free, optically controllable, and based on transfection of only two plasmids, i.e., an aav vector construct and one of six novel aav helper plasmids. the latter were engineered to carry aav serotype rep and cap genes together with adenoviral helper functions, as well as unique fluorescent p ... | 2003 | 12788658 |
| immune response following intraocular delivery of recombinant viral vectors. | there has been significant progress in the last few years in demonstrating the utility of recombinant viral vectors in treating a variety of ocular diseases. the field has moved beyond 'proof-of-principle' and, in fact, has entered the phase where some of these vectors/paradigms are being or soon will be evaluated in human clinical trials. for this reason and also, to increase the understanding of immunological effects of transgenes/viral vectors on the eye, it is important to summarize what is ... | 2003 | 12756418 |
| human gene targeting by adeno-associated virus vectors is enhanced by dna double-strand breaks. | the use of adeno-associated virus (aav) to package gene-targeting vectors as single-stranded linear molecules has led to significant improvements in mammalian gene-targeting frequencies. however, the molecular basis for the high targeting frequencies obtained is poorly understood, and there could be important mechanistic differences between aav-mediated gene targeting and conventional gene targeting with transfected double-stranded dna constructs. conventional gene targeting is thought to occur ... | 2003 | 12724413 |
| adeno-associated virus-mediated aspartoacylase gene transfer to the brain of knockout mouse for canavan disease. | canavan disease (cd) is an autosomal recessive leukodystrophy caused by deficiency of aspartoacylase (aspa). deficiency of aspa leads to elevation of n-acetyl-l-aspartic acid (naa) in the brain and urine. to explore the feasibility of gene transfer to replace aspa in cd, we generated a knockout mouse and constructed an aav vector that encodes human aspa cdna (haspa) followed by green fluorescent protein (gfp) after an intraribosomal entry site. we injected cd mice with raav-haspa-gfp in the stri ... | 2003 | 12718900 |
| 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 |
| [the in vitro isolation, culture and transfection of human fetal epidermal stem cells]. | to explore the in vitro methods of isolation and culture of human fetal epidermal stem cells (hfescs) and the feasibility of the cultured cells as the target cells for gene transfection. | 2003 | 12678969 |
| adeno-associated viral vector-mediated human vascular endothelial growth factor gene transfer stimulates angiogenesis and wound healing in the genetically diabetic mouse. | we studied the gene therapy efficacy of diabetes-associated wound healing disorder with an adeno-associated virus (aav) vector expressing the 165-amino acid isoform of human vascular endothelial growth factor-a (vegf-a) by using an incisional skin-wound model produced on the back of female diabetic c57bl/ksj db+/db+ mice and their normal littermates ( db+/+m). | 2003 | 12677400 |
| pka/prkx activity is a modulator of aav/adenovirus interaction. | interference between viruses occurs when infection by one virus results in the inhibition of replication of another virus. adeno-associated virus (aav2) is a human parvovirus with the unique characteristics of a dependence upon a helper virus for a productive infection and the ability to interfere with the replication of the helper virus. previously, we demonstrated that aav2 rep78 and rep52 interact and inhibit camp-dependent protein kinase a (pka) and its novel homolog prkx. we hypothesized th ... | 2003 | 12660177 |
| recombinant adeno-associated virus vectors efficiently and persistently transduce chondrocytes in normal and osteoarthritic human articular cartilage. | successful gene transfer into articular cartilage is a prerequisite for gene therapy of articular joint disorders. in the present study we tested the hypothesis that recombinant adeno-associated virus (raav) vectors are capable of effecting gene transfer in isolated articular chondrocytes in vitro, articular cartilage tissue in vitro, and sites of articular damage in vivo. using an raav vector carrying the escherichia coli beta-galactosidase gene (lacz) under the control of the cytomegalovirus ( ... | 2003 | 12659680 |
| efficient adeno-associated virus-mediated gene expression in human placenta-derived mesenchymal cells. | mesenchymal cells from various sources are pluripotent and are attractive sources for cell transplantation. in this study, we analyzed recombinant adeno-associated virus (raav)-mediated gene expression in human placenta-derived mesenchymal cells (hpdmcs), which reside in placental villi. after transduction of av-cag-egfp, a raav expressing enhanced green fluorescence protein (egfp), hpdmcs showed much higher level of egfp expression than human umbilical vein endothelial cells or rat aortic smoot ... | 2003 | 12636261 |
| production, purification and preliminary x-ray crystallographic studies of adeno-associated virus serotype 4. | adeno-associated virus (aav) serotypes 1 to 5 are currently under development as clinical gene delivery vectors for the treatment of human diseases. however, the ubiquitous nature of their cell surface receptors, heparin sulfate (aav2 and 3) and sialic acids (aav4 and 5), can preclude specific tissue targeting in vivo. structural studies of aav4 were initiated to characterize its capsid surface for re-targeting manipulations. crystals obtained diffracted synchrotron radiation to 3.2 a resolution ... | 2003 | 12620791 |
| nigrostriatal alpha-synucleinopathy induced by viral vector-mediated overexpression of human alpha-synuclein: a new primate model of parkinson's disease. | we used a high-titer recombinant adeno-associated virus (raav) vector to express wt or mutant human alpha-synuclein in the substantia nigra of adult marmosets. the alpha-synuclein protein was expressed in 90-95% of all nigral dopamine neurons and distributed by anterograde transport throughout their axonal and dendritic projections. the transduced neurons developed severe neuronal pathology, including alpha-synuclein-positive cytoplasmic inclusions and granular deposits; swollen, dystrophic, and ... | 2003 | 12601150 |
| the suppressor of cytokine signaling-1 (socs1) is a novel therapeutic target for enterovirus-induced cardiac injury. | enteroviral infections of the heart are among the most commonly identified causes of acute myocarditis in children and adults and have been implicated in dilated cardiomyopathy. although there is considerable information regarding the cellular immune response in myocarditis, little is known about innate signaling mechanisms within the infected cardiac myocyte that contribute to the host defense against viral infection. here we show the essential role of janus kinase (jak) signaling in cardiac my ... | 2003 | 12588885 |
| gene therapy delivery of endostatin enhances the treatment efficacy of radiation. | to evaluate whether sustained expression of mouse endostatin by adeno-associated virus (aav)-mediated gene transfer can enhance the treatment efficacy of ionizing radiation. | 2003 | 12559515 |
| [adeno-associated virus vector mediated gene transfer of hsvi-tk and its effect on killing cancer cell]. | the plasmid pactk-19 was constructed by inserting hsvi-tk gene into the multiple cloning sites of the general adeno-associated virus(aav) vector pacr-neo. when plasmid pactk-19 was transfected to recombinant aav's packaging cell line ae1201, which was exposed to adenovirus 5 for two hours before transfection, we got raav/actk at the titer of 3.4 x 10(5) cfu/ml. after infecting human lung cancer cell a549 with raav/actk, we extracted host cell's chromosome cdna and amplified part of the hsvi-tk s ... | 1998 | 12526317 |
| the kaposi's sarcoma-associated herpesvirus g protein-coupled receptor has broad signaling effects in primary effusion lymphoma cells. | kaposi's sarcoma-associated herpesvirus (kshv/human herpesvirus 8 [hhv-8]) is a gamma-2-herpesvirus responsible for kaposi's sarcoma as well as primary effusion lymphoma (pel). kshv is a lymphotropic virus that has pirated many mammalian genes involved in inflammation, cell cycle control, and angiogenesis. among these is the early lytic viral g protein-coupled receptor (vgpcr), a homologue of the human interleukin-8 (il-8) receptor. when expressed, vgpcr is constitutively active and can signal v ... | 2003 | 12477810 |
| dimerizer-regulated gene expression. | control of gene expression using small molecules is a powerful research tool and has clinical utility in the context of regulated gene therapy. use of chemical inducers of dimerization, or dimerizers, for this purpose has several advantages, including tight regulation, modularity to facilitate iterative improvements, and assembly from human proteins to minimize immune responses in clinical applications. recent developments include the use of the rapamycin-based dimerizer system to regulate the e ... | 2002 | 12459338 |
| conjugate-based targeting of recombinant adeno-associated virus type 2 vectors by using avidin-linked ligands. | the development of targeted vectors, capable of tissue-specific transduction, remains one of the important aspects of vector modification for gene therapy applications. recombinant adeno-associated virus type 2 (raav-2)-based vectors are nonpathogenic, have relatively low immunogenicity, and are capable of long-term transgene expression. aav-2 vectors bind primarily to heparan sulfate proteoglycan (hspg), a receptor that is present in many tissues and cell types. because of the widespread expres ... | 2002 | 12438615 |
| insect cells as a factory to produce adeno-associated virus type 2 vectors. | recombinant adeno-associated viruses (raav) are produced transiently in mammalian cells usually by cotransfecting two or three plasmids containing aav genes, adenovirus helper genes, and a vector genome. expansion and transfection of adherent cells limit the scale of raav production. efficient transfection is performed with cells on solid support media such as tissue culture plates. a large animal study or a human clinical trial may require 10(15) particles of vector, depending on dose. to gener ... | 2002 | 12427305 |
| vigilant vectors: adeno-associated virus with a biosensor to switch on amplified therapeutic genes in specific tissues in life-threatening diseases. | there are many life-threatening and chronic diseases in which physiological signals could be used to switch on therapeutic protective genes. we are developing a gene therapy approach in which a systemically injected "vigilant vector" waits for these signals and switches on genes to protect specific tissues with high amplification. the concept of a vigilant vector requires four components. the first component is a safe and stable vector that can be administered by systemic injection and express t ... | 2002 | 12413425 |
| production methods for gene transfer vectors based on adeno-associated virus serotypes. | vectors derived from adeno-associated virus serotype 2 (aav-2) represent a most promising tool for human gene transfer because these vectors are neither pathogenic nor toxic to the target cell, and allow long-term gene expression in a large variety of tissues. however, they are rather inefficient at infecting a number of clinically relevant cell types, and transduction by these vectors is likely hampered by neutralizing antibodies that are highly prevalent in the human population. therefore, an ... | 2002 | 12413413 |
| overexpression of parkinson's disease-associated alpha-synucleina53t by recombinant adeno-associated virus in mice does not increase the vulnerability of dopaminergic neurons to mptp. | mutations in the alpha-synuclein gene are linked to a rare dominant form of familial parkinson's disease, and alpha-synuclein is aggregated in lewy bodies of both sporadic and dominant parkinson's disease. it has been proposed that mutated alpha-synuclein causes dopaminergic neuron loss by enhancing the vulnerability of these neurons to a variety of insults, including oxidative stress, apoptotic stimuli, and selective dopaminergic neurotoxins, such as 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine ... | 2002 | 12360578 |
| primary human cells differ in their susceptibility to raav-2-mediated gene transfer and duration of reporter gene expression. | the susceptibility of a variety of different primary tissues was examined to long-term transduction with recombinant adeno-associated virus type 2 (raav-2) and factors influencing the transduction efficiency. in contrast to others using cell lines and animal models, emphasis was placed on the use of primary human cells. enhanced green fluorescent protein (egfp) marker gene expression was examined using fluorescence-activated cell sorting analysis. the most effective target cells for raav-2-media ... | 2002 | 12270659 |
| a p5 integration efficiency element mediates rep-dependent integration into aavs1 at chromosome 19. | adeno-associated virus (aav) undergoes site-specific integration into human chromosome 19 through a deletion-substitution mechanism at the well characterized aavs1 site. we have shown previously that a cis element within the left end of the aav genome enhances the efficiency of rep-mediated site-specific integration into chromosome 19 when present in inverted terminal repeat-containing recombinant aav (raav) plasmids. we now demonstrate that a 138-bp cis element, the p5 integration efficiency el ... | 2002 | 12221283 |
| light-activated gene transduction enhances adeno-associated virus vector-mediated gene expression in human articular chondrocytes. | to evaluate the effects of ultraviolet (uv) light as an adjuvant for recombinant adeno-associated virus (raav) transduction in human articular chondrocytes. | 2002 | 12209514 |
| transient platelet interaction induces mcp-1 production by endothelial cells via i kappa b kinase complex activation. | activated platelets alter endothelial chemotactic and adhesive properties. transient interaction of alpha-thrombin-activated platelets with endothelial cells is sufficient to induce secretion of the nf-kappa b-regulated chemokine mcp-1. to evaluate upstream signaling events in platelet-induced nf-kappa b activation, we compared the effect of platelets, il-1 beta or tnf-alpha on i kappa b kinase (ikk) complex activation and i kappa b phosphorylation/proteolysis. kinase assays demonstrated that pl ... | 2002 | 12195705 |
| neuroprotective effects of glial cell line-derived neurotrophic factor mediated by an adeno-associated virus vector in a transgenic animal model of amyotrophic lateral sclerosis. | amyotrophic lateral sclerosis (als) is a relentlessly progressive lethal disease that involves selective annihilation of motoneurons. glial cell line-derived neurotrophic factor (gdnf) is proposed to be a promising therapeutic agent for als and other motor neuron diseases. because adeno-associated virus (aav) has been developed as an attractive gene delivery system with proven safety, we explored the therapeutic efficacy of intramuscular delivery of the gdnf gene mediated by an aav vector (aav-g ... | 2002 | 12177190 |
| sustained high-level expression of human factor ix (hfix) after liver-targeted delivery of recombinant adeno-associated virus encoding the hfix gene in rhesus macaques. | the feasibility, safety, and efficacy of liver-directed gene transfer was evaluated in 5 male macaques (aged 2.5 to 6.5 years) by using a recombinant adeno-associated viral (raav) vector (raav-2 cagg-hfix) that had previously mediated persistent therapeutic expression of human factor ix (hfix; 6%-10% of physiologic levels) in murine models. a dose of 4 x 10(12) vector genomes (vgs)/kg of body weight was administered through the hepatic artery or portal vein. persistence of the raav vgs as circul ... | 2002 | 12176886 |
| generation of neutralizing activity against human immunodeficiency virus type 1 in serum by antibody gene transfer. | although several human immunodeficiency virus (hiv) vaccine approaches have elicited meaningful antigen-specific t-cell responses in animal models, no single vaccine candidate has engendered antibodies that broadly neutralize primary isolates of hiv type 1 (hiv-1). thus, there remains a significant gap in the design of hiv vaccines. to address this issue, we exploited the existence of rare human monoclonal antibodies that have been isolated from hiv-infected individuals. such antibodies neutrali ... | 2002 | 12163597 |
| a phase ii, double-blind, randomized, placebo-controlled clinical trial of tgaavcf using maxillary sinus delivery in patients with cystic fibrosis with antrostomies. | tgaavcf, an adeno-associated cystic fibrosis transmembrane conductance regulator (cftr) viral vector/gene construct, was administered to 23 patients in a phase ii, double-blind, randomized, placebo-controlled clinical trial. for each patient, a dose of 100,000 replication units of tgaavcf was administered to one maxillary sinus, while the contralateral maxillary sinus received a placebo treatment, thereby establishing an inpatient control. neither the primary efficacy endpoint, defined as the ra ... | 2002 | 12162817 |
| the atomic structure of adeno-associated virus (aav-2), a vector for human gene therapy. | the structure of the adeno-associated virus (aav-2) has been determined to 3-a resolution by x-ray crystallography. aav is being developed as a vector for gene therapy to treat diseases including hemophilia, cancer, and cystic fibrosis. as in the distantly related autonomous parvoviruses, the capsid protein has a beta-barrel fold, but long loops between the beta-strands share little structural homology with other parvoviruses, leading to unique surface features. most prominent are groups of thre ... | 2002 | 12136130 |
| tetracycline-inducible interleukin-10 gene transfer mediated by an adeno-associated virus: application to experimental arthritis. | the adeno-associated viruses (aav) offer new perspectives for cytokine gene transfer in rheumatoid arthritis (ra) because they are nonpathogenic and allow long-term transgene expression in vivo. moreover, the use of a tetracycline-inducible promoter allows regulation of therapeutic gene expression. this study assessed the potential long-term gene regulation of a recombinant aav vector expressing viral interleukin-10 (vil-10) in human rheumatoid synovium and the therapeutic efficiency in a mouse ... | 2002 | 12133271 |
| effects of adeno-associated virus-vectored ciliary neurotrophic factor on retinal structure and function in mice with a p216l rds/peripherin mutation. | past studies have shown that acute administration of ciliary neurotrophic factor (cntf) can prolong the survival of retinal photoreceptor cells that have undergone phototoxic injury or that express gene mutations. adenovirus-vectored cntf has also been effective but for all of these treatments, the effect has been transient. on the other hand, adeno-associated virus-vectored minigenes offer considerable promise for long-term survival. the authors sought to provide long-term, cntf-based protectio ... | 2002 | 12126945 |
| recombinant human parvovirus b19 vectors. | in an attempt to exploit the remarkable tissue-tropism of the human parvovirus b19 to target human hematopoietic cells of the erythroid lineage, recombinant human adeno-associated virus 2 genomes were encapsidated in parvovirus b19 capsids. although efficient transduction of primary human hematopoietic cells in the erythroid lineage occurred, a low-level of transgene expression in non-erythroid cells was also detected. these studies suggest that cell surface expression of p antigen, the primary ... | 2002 | 12116848 |
| second generation adeno-associated virus type 2-based gene therapy systems with the potential for preferential integration into aavs1. | adeno-associated virus type 2 (aav-2) is a non-pathogenic human parvovirus that is being developed as a gene therapy vector for the treatment of numerous diseases. one property of wild-type aav-2, that is highly desirable in a gene therapy vector, is its ability to preferentially integrate its dna into a 4 kilobase region of human chromosome 19, designated aavs1. one disadvantage of aav-2 is its relatively small packaging capacity, approximately 4.7 kilobases. because of this size limitation, th ... | 2002 | 12109212 |
| improved method of recombinant aav2 delivery for systemic targeted gene therapy. | a major hurdle in most current gene therapy modalities is the ability to transduce target tissues at very high efficiencies that ultimately lead to therapeutic levels of transgene expression. we have developed a novel method of recombinant adeno-associated virus 2 (raav) delivery that results in increased vector transduction efficiencies using microspheres reversibly conjugated to raav vectors. we hypothesize that conjugation to microspheres should result in a higher effective concentration of v ... | 2002 | 12095310 |
| targeted transgene insertion into human chromosomes by adeno-associated virus vectors. | efficient methods are needed for the precise genetic manipulation of diploid human cells, in which cellular senescence and low conventional gene targeting rates limit experimental and therapeutic options. we have shown previously that linear, single-stranded dna vectors based on adeno-associated virus (aav) could accurately introduce small (<20 bp) genetic modifications into homologous human chromosomal sequences. here we have used aav vectors to introduce large (>1 kb) functional transgene cass ... | 2002 | 12089561 |
| adeno-associated virus (aav)-mediated gene transfer in respiratory epithelium and submucosal gland cells in human fetal tracheal organ culture. | since the discovery of the cystic fibrosis transmembrane regulator (cftr) gene, cystic fibrosis has been an attractive target for gene therapy. postnatal gene transfer in the respiratory epithelium has been difficult and particularly inefficient in the submucosal gland cells, the target cells for cftr gene transfer. the authors hypothesized that during development, there is a favorable environment for fetal gene therapy with fewer physical barriers to efficient gene transfer and more accessible ... | 2002 | 12077770 |
| analysis of adeno-associated virus-mediated ex vivo transferred human beta-globin gene in bone marrow engrafted mice. | adeno-associated virus (aav)-2 was developed as a useful vector for human gene therapy. in this report, we analyzed the integration and expression of aav-mediated ex vivo transferred human beta-globin gene in bone marrow (bm) reconstituted mice. recombinant aav (raav) containing human beta-globin gene was packaged by infecting individual g418-resistant bhk-21 cell clones integrated with the plasmid av53hs432deltabeta2.0neo with recombinant herpes simplex virus, which can express rep and cap gene ... | 2002 | 12065900 |
| expression of human factor viii by splicing between dimerized aav vectors. | adeno-associated virus (aav) is a useful vector for hemophilia gene therapy, but the limited effective packaging capacity of aav (5 kb) appears to be incompatible with factor viii (gene symbol f8) cdna (7 kb). although we previously demonstrated efficient packaging and expression of b-domain-deleted human f8 (bdd-f8) using a single aav vector, the packaging limit still excludes the use of large/strong regulatory elements. here we exploited the split aav vector technology that expands the packagi ... | 2002 | 12027555 |
| adeno-associated virus protects the retinoblastoma family of proteins from adenoviral-induced functional inactivation. | adeno-associated virus type 2 (aav) is known to inhibit virally mediated oncogenic transformation. one of the early events of adenovirus (ad) infection is the functional inactivation of cell cycle regulatory retinoblastoma (rb) family of proteins, which consists of retinoblastoma protein (prb), p107, and p130. in an effort to understand the molecular basis of anti-oncogenic properties of aav, we studied the effects of aav expression on these proteins in cells infected with ad. western blot analy ... | 2002 | 12019182 |
| efficient integration of recombinant adeno-associated virus dna vectors requires a p5-rep sequence in cis. | the initial aim of this study was to combine attributes of adeno-associated virus (aav) and adenovirus (ad) gene therapy vectors to generate an ad-aav hybrid vector allowing efficient site-specific integration with ad vectors. in executing our experimental strategy, we found that, in addition to the known incompatibility of rep expression and ad growth, an equally large obstacle was presented by the inefficiency of the integration event when using traditional recombinant aav (raav) vectors. this ... | 2002 | 11991970 |
| rep/cap gene amplification and high-yield production of aav in an a549 cell line expressing rep/cap. | cell lines stably expressing rep/cap are important tools for studying adeno-associated virus (aav) biology and producing aav vectors. several rep/cap cell lines have been isolated, each of which is based on hela cells. infection of these cell lines with adenovirus for production of aav vector is associated with substantial amplification of the rep/cap gene. concerns over the presence of human papilloma viral (hpv) sequences in hela cells may limit use of such lines for production of clinical-gra ... | 2002 | 11991756 |
| adeno-associated virus capsids displaying immunoglobulin-binding domains permit antibody-mediated vector retargeting to specific cell surface receptors. | recombinant adeno-associated virus type 2 (raav2) is a promising vector for human somatic gene therapy. however, its broad host range is a disadvantage for some applications, because it reduces the specificity of the gene transfer. to overcome this limitation, we sought to create a versatile raav vector targeting system which would allow us to redirect raav binding to specific cell surface receptors by simple coupling of different ligands to its capsid. for this purpose, an immunoglobulin g (igg ... | 2002 | 11932421 |
| soluble flt-1 expression suppresses carcinomatous ascites in nude mice bearing ovarian cancer. | vascular endothelial growth factor (vegf), a bifunctional protein enhancing vascular permeability and stimulating endothelial growth, is thought to be responsible for fluid accumulation and angiogenesis in ascites tumors. to investigate the effects of stable expression of the soluble form of flt-1 vegf receptor (sflt-1), a known endogenous inhibitor of vegf, on the malignant ascites tumors, we cotransduced rmg-1 human ovarian cancer cells with adeno-associated virus vectors carrying the sflt-1 c ... | 2002 | 11929819 |
| recombinant adeno-associated virus gene therapy for cystic fibrosis and alpha(1)-antitrypsin deficiency. | 2002 | 11893723 | |
| [somatic gene therapy of dilated cardiomyopathy]. | the hereditary form of dilated cardiomyopathy (dcm) accounts for about 20% of human dcm and is a major cause of heart failure. to-2 strain hamsters show dcm, a gene deletion of delta-sarcoglycan (sg), loss of all four sgs, alpha-, beta-, gamma- and delta-sg proteins, and are useful for developing gene therapy of the hereditary dcm. the delta-sg is a component of dystrophin-associated glycoprotein complex that stabilizes sarcolemma. four familial and sporadic dcm cases have been reported in human ... | 2002 | 11862755 |
| recombinant adeno-associated virus serotype 2 vectors mediate stable interleukin 10 secretion from salivary glands into the bloodstream. | we have constructed a recombinant adeno-associated virus serotype 2 vector encoding human interleukin 10 (raavhil10). il-10 is a potent antiinflammatory/immune cytokine, which has received growing attention for its therapeutic potential. human il-10 (hil-10) production was virus dose dependent after in vitro infection of hsg cells, a human submandibular gland cell line. the vector-derived hil-10 produced was biologically active, as the medium from raavhil10-infected hsg cells caused a dose-depen ... | 2002 | 11812284 |
| rescue of hereditary form of dilated cardiomyopathy by raav-mediated somatic gene therapy: amelioration of morphological findings, sarcolemmal permeability, cardiac performances, and the prognosis of to-2 hamsters. | the hereditary form comprises approximately 1/5 of patients with dilated cardiomyopathy (dcm) and is a major cause of advanced heart failure. medical and socioeconomic settings require novel treatments other than cardiac transplantation. to-2 strain hamsters with congenital dcm show similar clinical and genetic backgrounds to human cases that have defects in the delta-sarcoglycan (delta-sg) gene. to examine the long-term in vivo supplement of normal delta-sg gene driven by cytomegalovirus promot ... | 2002 | 11805334 |
| neurological correction of lysosomal storage in a mucopolysaccharidosis iiib mouse model by adeno-associated virus-mediated gene delivery. | mucopolysaccharidosis (mps) iiib is characterized by mild somatic features and severe neurological diseases leading to premature death. no definite treatment is available for mps iiib patients. we constructed two recombinant adeno-associated virus (raav) vectors containing the human alpha-n-acetylglucosaminidase (naglu) cdna driven by either a cmv or a neuron-specific enolase (nse) promoter. in vitro, these raav vectors mediated efficient expression of recombinant naglu in human mps iiib fibrobl ... | 2002 | 11786044 |
| rapid induction of cytotoxic t-cell response against cervical cancer cells by human papillomavirus type 16 e6 antigen gene delivery into human dendritic cells by an adeno-associated virus vector. | we have shown that the pulsing of dendritic cells (dcs) with human papillomavirus type 16 (hpv-16) antigen proteins by lipofection stimulates class i-restricted cytotoxic t lymphocyte (ctl) response against primary cervical cancer cells. also, we have shown that adeno-associated virus (aav) was able to effectively deliver a cytokine gene into dcs. it has been our hypothesis that the delivery of antigen genes into dcs, resulting in endogenous and continuous antigen protein expression, may result ... | 2001 | 11781657 |
| a high-capacity, capsid-modified hybrid adenovirus/adeno-associated virus vector for stable transduction of human hematopoietic cells. | to achieve stable gene transfer into human hematopoietic cells, we constructed a new vector, deltaad5/35.aav. this vector has a chimeric capsid containing adenovirus type 35 fibers, which conferred efficient infection of human hematopoietic cells. the deltaad5/35.aav vector genome is deleted for all viral genes, allowing for infection without virus-associated toxicity. to generate high-capacity deltaad5/35.aav vectors, we employed a new technique based on recombination between two first-generati ... | 2002 | 11773389 |
| exchange of surface proteins impacts on viral vector cellular specificity and transduction characteristics: the retina as a model. | recombinant vectors based on adeno-associated virus (aav) or human immunodeficiency 1 (lentivirus) are promising tools for long term in vivo gene delivery. their design allows the exchange of capsids or envelopes, respectively, theoretically providing the opportunity to transduce a range of cell types. we constructed aav vectors encoding enhanced green fluorescent protein (egfp) within an aav serotype 2 (aav2) genome contained in an aav2, five or one capsid (called aav2/2, aav2/5 and aav2/1, res ... | 2001 | 11751689 |
| glial cell line derived neurotrophic factor delays photoreceptor degeneration in a transgenic rat model of retinitis pigmentosa. | we designed experiments to evaluate the therapeutic potential of glial cell line derived neurotrophic factor (gdnf) to rescue photoreceptors from genetically determined cell death. gene transfer of the neurotrophic factor to the retina was achieved via a recombinant adeno-associated virus (raav) vector containing the chicken beta-actin promoter/immediate early cytomegalovirus enhancer (cba) driving the human gdnf gene. we delivered aav-cba-gdnf to the retinas of an animal model of retinitis pigm ... | 2001 | 11735347 |
| 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 |
| 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 |
| 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 |
| 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 |
| adeno-associated virus-mediated delivery of glial cell line-derived neurotrophic factor protects motor neuron-like cells from apoptosis. | motor neuron disorders including amyotrophic lateral sclerosis may benefit from the induction of neurotrophic factors such as glial cell line-derived neurotrophic factor (gdnf) that are known to be trophic and protective for motor neurons. however, the application of such factors is limited by an inability to successfully target their expression in the nervous system. in this study we investigate the potential of using adeno-associated virus (aav) as a vector for gene delivery into motor neuron- ... | 2001 | 11582516 |
| stable therapeutic serum levels of human alpha-1 antitrypsin (aat) after portal vein injection of recombinant adeno-associated virus (raav) vectors. | previous work from our group showed that recombinant adeno-associated virus (raav) vectors mediated long-term secretion of therapeutic serum levels of human alpha-1 antitrypsin (haat) after a single injection in murine muscle. we hypothesized that hepatocyte transduction could be even more efficient, since these cells represent the natural site of aat production and secretion. to test this hypothesis, raav vectors containing the haat cdna driven by either the human elongation factor 1 alpha prom ... | 2001 | 11571566 |
| insertional mutagenesis of the adeno-associated virus type 2 (aav2) capsid gene and generation of aav2 vectors targeted to alternative cell-surface receptors. | recombinant adeno-associated virus (aav) vectors are of interest in the context of gene therapy because of their ability to mediate efficient transfer and stable expression of therapeutic genes in a wide variety of tissues. however, aav-mediated gene delivery to specific cell populations is often precluded by the widespread distribution of heparan sulfate proteoglycan (hspg), the primary cellular receptor for the virus. conversely, an increasing number of cell types are being identified that do ... | 2001 | 11560765 |
| adeno-associated virus type 2-mediated gene transfer: role of cellular fkbp52 protein in transgene expression. | although adeno-associated virus type 2 (aav) has gained attention as a potentially useful vector for human gene therapy, the transduction efficiencies of aav vectors vary greatly in different cells and tissues in vitro and in vivo. we have documented that a cellular tyrosine phosphoprotein, designated the single-stranded d-sequence-binding protein (ssd-bp), plays a crucial role in aav-mediated transgene expression (k. y. qing, x.-s. wang, d. m. kube, s. ponnazhagan, a. bajpai, and a. srivastava, ... | 2001 | 11533160 |
| characterization of permanent cell lines that contain the aav2 rep-cap genes on an epstein-barr-virus-based episomal plasmid. | recombinant adeno-associated virus (raav) has emerged as a promising gene therapy vector. its development, however, has been hampered by the lack of a readily available efficient production method. we investigated the possibility of establishing permanent cell lines for the production of raav with a new epstein-barr-virus (ebv)-based episomal aav rep-cap plasmid (pcep-rep/cap). hela and 293 cells were stably transfected with plasmids that carry the aav2 rep/cap genes under transcriptional contro ... | 2001 | 11509889 |
| enhancement of muscle gene delivery with pseudotyped adeno-associated virus type 5 correlates with myoblast differentiation. | adeno-associated virus (aav)-based muscle gene therapy has achieved tremendous success in numerous animal models of human diseases. recent clinical trials with this vector have also demonstrated great promise. however, to achieve therapeutic benefit in patients, large inocula of virus will likely be necessary to establish the required level of transgene expression. for these reasons, efforts aimed at increasing the efficacy of aav-mediated gene delivery to muscle have the potential for improving ... | 2001 | 11462038 |
| incorporation of tumor-targeting peptides into recombinant adeno-associated virus capsids. | the human parvovirus adeno-associated virus type 2 (aav-2) possesses many features that make it an attractive vector for gene delivery in vivo. however, its broad host range may limit its usefulness and effectivity in several gene therapy applications in which transgene expression needs to be limited to a specific organ or cell type. in this study, we explored the possibility of directing recombinant aav-2 transduction by incorporating targeting peptides previously isolated by in vivo phage disp ... | 2001 | 11407911 |
| adeno-associated virus 2-mediated transduction and erythroid lineage-restricted long-term expression of the human beta-globin gene in hematopoietic cells from homozygous beta-thalassemic mice. | adeno-associated virus 2 (aav), a nonpathogenic human parvovirus, has gained attention as a potentially useful vector for human gene therapy. here, we report successful aav-mediated stable transduction and high-efficiency, long-term, erythroid lineage-restricted expression of a human beta-globin gene in primary murine hematopoietic stem cells in vivo. bone marrow-derived primitive sca-1(+), lin(-) hematopoietic stem cells from homozygous beta-thalassemic mice were transduced ex vivo with a recom ... | 2001 | 11407908 |
| hybrid vectors based on adeno-associated virus serotypes 2 and 5 for muscle-directed gene transfer. | vectors based on hybrids consisting of adeno-associated virus types 2 (itrs and rep) and 5 (cap) were evaluated for muscle-directed gene transfer (called aav2/5). evaluation in immune-competent mice revealed greater transduction efficacy with aav2/5 than with aav2 and no cross-neutralization between aav2/5 and aav2. interestingly, we saw no immunologic evidence of previous exposure to aav5 capsids in a large population of healthy human subjects. | 2001 | 11390622 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column. | one of the most promising gene transfer vectors in human clinical trials is aav2. the quality of the vector preparations is a key element in obtaining reliable and reproducible data in preclinical studies. however, established protocols either result in impure, low infectious virus (cscl2 gradient centrifugation) or demand a high level of manual and technical skills (cscl2 gradient centrifugation, iodixanol/heparin or hplc purification). in this study, we present an easy-to-do single-step column ... | 2001 | 11177544 |
| intracellular trafficking of adeno-associated virus vectors: routing to the late endosomal compartment and proteasome degradation. | the early steps of adeno-associated virus (aav) infection involve attachment to a variety of cell surface receptors (heparan sulfate, integrins, and fibroblast growth factor receptor 1) followed by clathrin-dependent or independent internalization. here we have studied the subsequent intracellular trafficking of aav particles from the endosomal compartment to the nucleus. human cell lines were transduced with a recombinant aav (raav) carrying a reporter gene (luciferase or green fluorescent prot ... | 2001 | 11160681 |
| 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 |
| 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 |
| 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 |
| 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 type 2 rep78 induces apoptosis through caspase activation independently of p53. | adeno-associated virus (aav) type 2 rep78 is a multifunctional protein required for aav dna replication, integration, and gene regulation. the biochemical activities of rep78 have been described, but the effects of rep proteins on the cell have not been characterized. we have analyzed rep-mediated cytotoxicity. we demonstrated that rep78 expression is sufficient to induce cell death and disruption of the cell cycle. cell death was found to be mediated by apoptosis. rep78 expression resulted in t ... | 2000 | 11000213 |
| use of the nadh-quinone oxidoreductase (ndi1) gene of saccharomyces cerevisiae as a possible cure for complex i defects in human cells. | the ndi1 enzyme of saccharomyces cerevisiae is a single subunit rotenone-insensitive nadh-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. we have shown previously that the ndi1 gene can be functionally expressed in chinese hamster cells (seo, b. b., kitajima-ihara, t., chan, e. k., scheffler, i. e., matsuno-yagi, a., and yagi, t. (1998) proc. natl. acad. sci. u. s. a. 95, 9167-9171) and human embryonal kidney 293 (hek 293) cells (seo, b. b., matsuno ... | 2000 | 10982813 |
| selective cleavage of aavs1 substrates by the adeno-associated virus type 2 rep68 protein is dependent on topological and sequence constraints. | the adeno-associated virus type 2 (aav-2) rep78 and rep68 proteins are required for replication of the virus as well as its site-specific integration into a unique site, called aavs1, of human chromosome 19. rep78 and rep68 initiate replication by binding to a rep binding site (rbs) contained in the aav-2 inverted terminal repeats (itrs) and then specifically nicking at a nearby site called the terminal resolution site (trs). similarly, rep78 and rep68 are postulated to trigger the integration p ... | 2000 | 10982325 |
| hyaluronidase enhances recombinant adeno-associated virus (raav)-mediated gene transfer in the rat skeletal muscle. | skeletal muscle is a privileged target for long-term raav-mediated gene transfer in mouse, rat, dog and non-human primates. intramuscular injections of raav encoding human factor ix in hemophilia b patients have been initiated, based on promising results gathered in affected dogs. we found that intramuscular raav administration in rats resulted in restricted transduction essentially along the myofibers axis with poor lateral diffusion. this suggested that the transduction rate might be limited b ... | 2000 | 10981669 |
| inhibition of s-phase progression by adeno-associated virus rep78 protein is mediated by hypophosphorylated prb. | adeno-associated virus (aav) has an antiproliferative action on cells. we investigated the effect of the aav replication proteins (rep) on the cell division cycle using retroviral vectors. rep78 and rep68 inhibited the growth of primary, immortalized and transformed cells, while rep52 and rep40 did not. rep68 induced cell cycle arrest in phases g(1) and g(2), with elevated cdk inhibitor p21 and reduced cyclin e-, a- and b1-associated kinase activity. rep78-expressing cells were also impaired in ... | 2000 | 10944118 |
| 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 |
| increasing the size of raav-mediated expression cassettes in vivo by intermolecular joining of two complementary vectors. | a major shortcoming to the use of adeno-associated virus (raav) vectors is their limited packaging size. to overcome this hurdle, we split an expression cassette and cloned it into two separate vectors. the vectors contained either a nuclear localizing escherichia coli lacz transgene (nlslacz) with a splice acceptor, or the human elongation factor 1alpha ( ef1alpha) gene enhancer/promoter(s) (ef1alphaep) with a splice donor. we co-injected a promoter-less nlslacz vector with a vector containing ... | 2000 | 10802620 |
| gene delivery to human chondrocytes by an adeno associated virus vector. | to investigate the efficiency of gene transduction to human chondrocytes using an adeno associated virus (aav) vector. | 2000 | 10782826 |
| design and packaging of adeno-associated virus gene targeting vectors. | adeno-associated virus (aav) vectors can transduce cells by several mechanisms, including (i) gene addition by chromosomal integration or episomal transgene expression or (ii) gene targeting by modification of homologous chromosomal sequences. the latter process can be used to correct a variety of mutations in chromosomal genes with high fidelity and specificity. in this study, we used retroviral vectors to introduce mutant alkaline phosphatase reporter genes into normal human cells and subseque ... | 2000 | 10775597 |