Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
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| recombinant aav viral vectors pseudotyped with viral capsids from serotypes 1, 2, and 5 display differential efficiency and cell tropism after delivery to different regions of the central nervous system. | recombinant adeno-associated virus 2 (raav2) has been shown to deliver genes to neurons effectively in the brain, retina, and spinal cord. the characterization of new aav serotypes has revealed that they have different patterns of transduction in diverse tissues. we have investigated the tropism and transduction frequency in the central nervous system (cns) of three different raav vector serotypes. the vectors contained aav2 terminal repeats flanking a green fluorescent protein expression casset ... | 2004 | 15294177 |
| infectious titer assay for adeno-associated virus vectors with sensitivity sufficient to detect single infectious events. | a highly sensitive assay for determination of infectious titers of recombinant adeno-associated virus (aav) by limiting dilution analysis is described. this assay is capable of detecting single infectious events and can therefore provide an absolute rather than relative measure of infectivity. the assay utilizes a hela-derived aav2 rep/cap-expressing cell line, d7-4, grown in 96-well plates and infected with replicate 10-fold serial dilutions of aav2 vectors in the presence of adenovirus type 5. ... | 2004 | 15298029 |
| restriction of neuroblastoma angiogenesis and growth by interferon-alpha/beta. | we tested the hypothesis that the antiangiogenic activity of the type i interferons (ifns), could affect tumor engraftment and growth in murine xenograft models of neuroblastoma. | 2004 | 15300178 |
| attenuation of dengue virus infection by adeno-associated virus-mediated sirna delivery. | background: the need for safe and effective treatment of dengue virus (den), a class a agent that causes dengue hemorrhagic fever/dengue shock syndrome, has been a critical global priority. an effective vaccine for den is not yet available. in this study the possibility of attenuating den infection using adeno-associated virus (aav)-encoded short interfering rnas (sirna) was examined in vero cells and human dendritic cells (dcs). methods: a cassette encoding sirna targeted to a 3' untranslated s ... | 2004 | 15301687 |
| structure of adeno-associated virus type 2 rep40-adp complex: insight into nucleotide recognition and catalysis by superfamily 3 helicases. | we have determined the structure of adeno-associated virus type 2 (aav2) rep40 to 2.1-a resolution with adp bound at the active site. the complex crystallizes as a monomer with one adp molecule positioned in an unexpectedly open binding site. the nucleotide-binding pocket consists of the p-loop residues interacting with the phosphates and a loop (nucleoside-binding loop) that emanates from the last strand of the central beta-sheet and interacts with the sugar and base. as a result of the open na ... | 2004 | 15310852 |
| expression of vascular endothelial growth factor (vegf) in human osteosarcoma cells transfected with adeno-associated virus-antisense vegf. | the expression of protein vascular endothelial growth factor (vegf) in osteosarcoma cells transfected with adeno-associated virus (raav)-antisense vegf was studied to provide the foundation of osteosarcoma treatment through antivascularization. the raav-antisense vegf at different doses (0, 20, 50, 100, 200, 240 microl) was transfected into osteosarcoma mg-63 cell. the cells and culture supernatants were collected before and after tansfection. the expression of vegf protein was detected by using ... | 2004 | 15315348 |
| therapeutic levels of factor ix expression using a muscle-specific promoter and adeno-associated virus serotype 1 vector. | extensive studies in animal models of the x-linked bleeding disorder hemophilia b (deficiency in functional coagulation factor ix, f.ix) have shown that muscle-directed adeno-associated (aav)-mediated f.ix gene transfer can be used to treat this disease. however, large vector doses of aav-2 vector are required for therapeutic levels of expression, and the number of vector doses that can be injected per intramuscular site is limited. several studies have shown that some of these limitations can b ... | 2004 | 15319035 |
| hiv-1 integration sites are flanked by potential mars that alone can act as promoters. | matrix attachment regions (mars) are cis regulatory elements that modulate gene expression in a tissue and cell stage specific manner. recent reports show that viral integration within the genome takes place at nonrandom active genes. we have checked for the presence of mars in the vicinity of the reported 524 hiv-1 integration sites. our studies show that in 92.5% cases, mars flank the integration sites. similarly, for adeno-associated virus, two potential mars were present next to the integrat ... | 2004 | 15325282 |
| widespread correction of lysosomal storage following intrahepatic injection of a recombinant adeno-associated virus in the adult mps vii mouse. | mucopolysaccharidosis type vii is a lysosomal storage disease caused by deficiency of the acid hydrolase beta-glucuronidase. mps vii mice develop progressive lysosomal accumulation of glycosaminoglycans within multiple organs, including the brain. using this animal model, we investigated whether gene transfer mediated by a recombinant adeno-associated virus (raav) type 2 vector is capable of reversing the progression of storage in adult mice. we engineered an raav2 vector to carry the murine bet ... | 2004 | 15336648 |
| differential myocardial gene delivery by recombinant serotype-specific adeno-associated viral vectors. | recombinant cross-packaging of adeno-associated virus (aav) genome of one serotype into other aav serotypes has the potential to optimize tissue-specific gene transduction and expression in the heart. to evaluate the role of aav1 to 5 virion shells on aav2 transgene transduction, we constructed hybrid vectors in which each serotype capsid coding domain was cloned into a common vector backbone containing aav2 replication genes. constructs were tested for expression in: (1) adult murine heart in v ... | 2004 | 15336660 |
| targeted gene delivery to vascular tissue in vivo by tropism-modified adeno-associated virus vectors. | gene therapy offers an unprecedented opportunity to treat diverse pathologies. adeno-associated virus (aav) is a promising gene delivery vector for cardiovascular disease. however, aav transduces the liver after systemic administration, reducing its usefulness for therapies targeted at other sites. because vascular endothelial cells (ecs) are in contact with the bloodstream and are heterogeneous between organs, they represent an ideal target for site-specific delivery of biological agents. | 2004 | 14732747 |
| immunity to adeno-associated virus serotype 2 delivered transgenes imparted by genetic predisposition to autoimmunity. | adeno-associated virus (aav) is widely considered a promising vector for therapeutic gene delivery. this promise is based on previous studies assessing aavs safety and toxicity, ability to infect nondividing cells, elicit a limited immune response and provide long-term gene expression. however, we now find that earlier studies underappreciated the degree of aav immunogenicity as well as the extent to which genetic background, through regulation of immune responsiveness, influences the duration o ... | 2004 | 14737082 |
| autoimmune anemia in macaques following erythropoietin gene therapy. | we delivered the homologous erythropoietin (epo) cdna driven from a doxycycline-regulated promoter via recombinant adeno-associated virus in skeletal muscle of 9 cynomolgus macaques. upon induction, rapid supraphysiologic levels of epo were obtained. unexpectedly, some individuals developed a profound anemia that correlated with the appearance of neutralizing antibodies against the endogenous epo. both the endogenous erythropoietin and vector sequences were identical. this is the first example o ... | 2004 | 14739218 |
| evidence of chromosomal integration of aav dna in human testis tissue. | persistent infection with adeno-associated virus (aav) has been demonstrated in human tissues, most frequently in the female and male genital tract. the clinical significance of latent aav infection remains, however, uncertain to date. the mode of latency of aav is not known, i.e., it is unclear whether the viral genome is integrated in the cellular genome, and if integration occurs site-specifically in chromosome 19 as has been observed in cell culture. therefore we investigated if viral dna in ... | 2004 | 14739652 |
| anti-tumor efficacy of human angiostatin using liver-mediated adeno-associated virus gene therapy. | angiostatin is a potent endogenous inhibitor of angiogenesis and tumor growth in vivo. the therapeutic potential of adeno-associated viral (aav) gene delivery of angiostatin in modulating tumor growth in vivo was evaluated. sustained levels of angiostatin were detected in the sera of mice for up to 6 months after they received a single injection of aav-angiostatin. aav-mediated stable expression of angiostatin inhibited tumor burden in the highly aggressive b16f10 melanoma and lewis lung carcino ... | 2004 | 14741778 |
| regulation of a ca2+-activated k+ channel by calcium-sensing receptor involves p38 map kinase. | by using pharmacological and molecular approaches, we previously showed that the g-protein-coupled, extracellular calcium (ca2+(o))-sensing receptor (car) regulates a large-conductance (approximately 140 ps), ca(2+)-activated k+ channel [ik(ca); cakc] in u87 astrocytoma cells. here we show that elevated ca2+(o) stimulates extracellular-signal-regulated kinase (erk1/2) and p38 map kinase (mapk). the effect of high ca2+(o) on p38 mapk but not erk1/2 is car mediated, insofar as transduction with a ... | 2004 | 14743432 |
| alternative polyadenylation of adeno-associated virus type 5 rna within an internal intron is governed by the distance between the promoter and the intron and is inhibited by u1 small nuclear rnp binding to the intervening donor. | adeno-associated virus type 5 is unique among adeno-associated virus serotypes in that it uses a polyadenylation site in the center of the genome. the great majority of transcripts generated from the upstream p7 and p19 promoters are polyadenylated at a site in the central intron ((pa)p); however, most of the viral transcripts generated by the proximal p41 promoter are polyadenylated at the distal polyadenylation site at the 3' end of the genome (pa)d and subsequently spliced. polyadenylation at ... | 2004 | 14749332 |
| gene therapy for leber congenital amaurosis. | recent success in delivering vision to a canine model of a severe, early-onset blinding disease, leber congenital amaurosis (lca) (acland et al 2001) demonstrates that adeno-associated virus serotype 2 (aav2) is capable of delivering a corrective gene to the target retinal cells. results of these studies indicate long-term rescue of vision as assessed by psychophysical, behavioural and molecular biological studies. preliminary results of studies in progress are described and the implications of ... | 2004 | 14750605 |
| a new animal model for pulmonary hypertension based on the overexpression of a single gene, angiopoietin-1. | angiopoietin-1 gene expression in human pulmonary hypertensive lungs is directly proportional to increasing pulmonary vascular resistance. we hypothesized that targeted overexpresssion of angiopoietin-1 in the lung would cause persistent pulmonary hypertension in an animal model. | 2004 | 14759414 |
| in utero gene therapy rescues vision in a murine model of congenital blindness. | the congenital retinal blindness known as leber congenital amaurosis (lca) can be caused by mutations in the rpe65 gene. rpe65 plays a critical role in the visual cycle that produces the photosensitive pigment rhodopsin. recent evidence from human studies of lca indicates that earlier rather than later intervention may be more likely to restore vision. we determined the impact of in utero delivery of the human rpe65 cdna to retinal pigment epithelium cells in a murine model of lca, the rpe65(-/- ... | 2004 | 14759802 |
| development of efficient viral vectors selective for vascular smooth muscle cells. | the vascular smooth muscle cell (smc) is integral to the pathogenesis of neointimal formation associated with late vein graft failure, in-stent restenosis, and transplant arteriopathy. viral vectors transduce smc with low efficiency and hence, there is a need for improvement. we aimed to enhance the efficiency and selectivity of gene delivery to human smc. targeting ligands were identified using phage display on primary human saphenous vein smc with linear and cyclic libraries. two linear peptid ... | 2004 | 14759804 |
| the effects of raav2-mediated ngf gene delivery in adult and aged rats. | nerve growth factor (ngf) therapy has been proposed to treat patients with age-related cognitive deficits, including those with alzheimer's disease. one promising approach to delivering this protein into brain involves viral vectors. however, little is known about the effects of aging on gene transfer in brain generally and in particular its effect on transgenic ngf expression. to examine the transgene expression and biological effects of ngf gene transfer in adult and aged rats, we delivered mo ... | 2004 | 14759810 |
| dna-dependent pk inhibits adeno-associated virus dna integration. | recent studies have shown that recombinant adeno-associated virus (raav) can persist in episomal form; however, factors affecting raav persistence are poorly understood. dna-dependent pk (dna-pk) is a dna repair enzyme, which we previously found played an important role in determining the molecular fate of the raav genome in mouse skeletal muscle. in the present study, we tested the effect of dna-pk on aav serotype 2 integration in vitro and in vivo in mouse liver. in an in vitro integration sys ... | 2004 | 14766968 |
| effect of hydroxyurea and etoposide on transduction of human bone marrow mesenchymal stem and progenitor cell by adeno-associated virus vectors. | to study the effect of hydroxyurea and etoposide on transduction of human marrow mesenchymal and progenitor stem cells by adeno-associated virus (aav). | 2004 | 14769209 |
| designing gene delivery vectors for cardiovascular gene therapy. | genetic therapy in the cardiovascular system has been proposed for a variety of diseases ranging from prevention of vein graft failure to hypertension. such diversity in pathogenesis requires the delivery of therapeutic genes to diverse cell types in vivo for varying lengths of time if efficient clinical therapies are to be developed. data from extensive preclinical studies have been compiled and a certain areas have seen translation into large-scale clinical trials, with some encouraging report ... | 2004 | 14769440 |
| repeated adeno-associated virus serotype 2 aerosol-mediated cystic fibrosis transmembrane regulator gene transfer to the lungs of patients with cystic fibrosis: a multicenter, double-blind, placebo-controlled trial. | the primary objective was to determine the safety and tolerability of repeated doses of aerosolized adeno-associated serotype 2 vector containing cystic fibrosis transmembrane conductance regulator (cftr) complementary dna (cdna) [tgaavcf], an adeno-associated virus (aav) vector encoding the complete human cftr cdna. secondary objectives included evaluation of pulmonary function assessed by spirometry, lung abnormalities by high-resolution ct (hrct), airway cytokines, vector shedding, serum neut ... | 2004 | 14769732 |
| gdnf gene therapy attenuates retinal ischemic injuries in rats. | to examine the protective effects of glial cell line-derived neurotrophic factor (gdnf) on retinal ischemia-reperfusion injury by using gene delivery. | 2004 | 14961006 |
| transgene expression after rep-mediated site-specific integration into chromosome 19. | we have used a plasmid-based transfection model of the adeno-associated virus (aav) rep-mediated site-specific integration (rmssi) pathway to characterize the stability and expression of a site-specifically integrated transgene (either green fluorescent protein [gfp] or chloramphenicol acetyltransferase [cat]). three plasmids containing the aav p5 integration efficiency element (p5iee) have been used to study integration and transgene expression in hela cells: (1) prepgfp(itr+) contains both aav ... | 2004 | 14965377 |
| phase i trial of intramuscular injection of a recombinant adeno-associated virus alpha 1-antitrypsin (raav2-cb-haat) gene vector to aat-deficient adults. | a recombinant virus vector constructed from adeno-associated virus (aav) that has been altered to carry the human alpha1-antitrypsin (haat) gene expressed from a hybrid chicken beta-actin promoter with a cytomegalovirus enhancer has been developed. the construct has been shown to initiate the production of haat in animal models closely matching the proposed human trial. the proposed clinical trial is an open-label, phase i study administering recombinant adeno-associated virus alpha1-antitrypsin ... | 2004 | 14965381 |
| the nuclease domain of adeno-associated virus rep coordinates replication initiation using two distinct dna recognition interfaces. | integration into a particular location in human chromosomes is a unique property of the adeno-associated virus (aav). this reaction requires the viral rep protein and aav origin sequences. to understand how rep recognizes dna, we have determined the structures of the rep endonuclease domain separately complexed with two dna substrates: the rep binding site within the viral inverted terminal repeat and one of the terminal hairpin arms. at the rep binding site, five rep monomers bind five tetranuc ... | 2004 | 14967147 |
| overview of adeno-associated viral vectors. | the use of adeno-associated virus (aav) as a gene transfer vector has been steadily increasing over the past several years. aav vectors have been particularly useful for applications where sustained gene expression is required. prolonged in vivo expression following aav treatment has been seen in the liver (1,2), brain (3,4), skeletal muscle (5,6), lung (7,8), and hematopoietic stem cells (9,10) of animal models. therapeutic benefit from aav treatment has been shown in a number of preclinical mo ... | 2004 | 14970590 |
| aav vector delivery to cells in culture. | adeno-associated virus (aav) gene delivery vectors are being investigated as vehicles for gene therapy for a wide variety of hereditary and acquired human diseases. aav's inability to self-propagate, ability to be maintained as an episome in the transduced cell, and relatively innocuous effects on the immune system make it the vector of choice for prolonged in vivo gene expression. aav type 2 is the most commonly used serotype for gene delivery. aav2 vectors will deliver dna to a wide variety of ... | 2004 | 14970591 |
| aav-mediated gene transfer to the liver. | the liver is a frequent target of gene-transfer experiments, because of its central role in many metabolic and synthetic pathways. for applications where prolonged expression of genes in the liver is required, adeno-associated virus (aav) has proven to be an effective tool for in vivo gene transfer. high-level, persistent hepatic expression has been achieved in a number of experimental systems following a single treatment with aav in murine and larger animal models. this prolonged expression is ... | 2004 | 14970593 |
| gene delivery to the mammalian heart using aav vectors. | there are a large number of cardiovascular diseases that could be treated by myocardial gene transfer. these include congestive heart failure, ischemic heart disease, and cardiomyopathy. in addition to its potential for treatment of disease, myocardial gene transfer is useful for the analysis of gene expression and promoter function and for generating animal models of human disease such as pulmonary hypertension. the ideal vector for myocardial gene therapy should give efficient and stable trans ... | 2004 | 14970595 |
| gene delivery to the mouse brain with adeno-associated virus. | the efficient transduction of postmitotic cells by adeno-associated virus (aav) makes it an excellent vector to deliver marker, functional, or therapeutic genes to the mammalian brain. an attractive feature of aav is that all the viral-coding sequences are removed when engineering the recombinant genome, thereby limiting the extent of cell toxicity and immune response that are often associated with viral gene transcription. of the seven described aav serotypes, aav serotype-2 (aav2) is the most ... | 2004 | 14970596 |
| delivery of dna to tumor cells in vivo using adeno-associated virus. | the number of published studies on transduction of tumor cells in vivo using recombinant adeno-associated virus (raav) vectors is very limited compared with those that have been published on targeting normal cells. a major reason for this can be attributed to the biology of the vector itself. aav, being a nonpathogenic vector capable of providing transgene integration and long-term expression, is ideally suited for the correction of metabolic defects either to replace a defective protein/enzyme ... | 2004 | 14970597 |
| gene delivery to human and murine primitive hematopoietic stem and progenitor cells by aav2 vectors. | the adeno-associated virus 2 (aav2) is known to possess a broad host-range that transcends the species barrier. the broad host-range and nonpathogenic nature of aav, coupled with its site-specificity and stable integration of the proviral genome, have led to the development of recombinant aav vectors (2). recombinant aav vectors have been shown to transduce certain cell types, such as muscle and brain, exceedingly well (3-5). however, controversies exist with regard to the efficacy of aav vector ... | 2004 | 14970598 |
| delivery of genes to the eye using lentiviral vectors. | the primary aim of gene transfer into the retinal cells has been to investigate the developmental mechanisms of the retinal cells or to reverse retinal diseases. retroviruses have been used to investigate the differentiation of retinal cells, to study the embryonic retina in vivo or explant organ culture, and to trace the fate of the cells that were dividing at the time of gene transfer. using adenovirus, bennett et al. showed the possibility of using gene therapy to correct degenerative disease ... | 2004 | 14970609 |
| gene targeting in stem cells from individuals with osteogenesis imperfecta. | adult stem cells offer the potential to treat many diseases through a combination of ex vivo genetic manipulation and autologous transplantation. mesenchymal stem cells (mscs, also referred to as marrow stromal cells) are adult stem cells that can be isolated as proliferating, adherent cells from bones. mscs can differentiate into multiple cell types present in several tissues, including bone, fat, cartilage, and muscle, making them ideal candidates for a variety of cell-based therapies. here, w ... | 2004 | 14976317 |
| adeno-associated viral vector-mediated apoe expression in alzheimer's disease mice: low cns immune response, long-term expression, and astrocyte specificity. | recombinant adenovirus and adeno-associated virus (aav) are highly effective vehicles for gene transfer into cns cells. however, the duration of gene expression and the cytotoxicity to cells are quite different between these viral approaches. we initially investigated these distinctions by stereotaxically injecting both adenovirus vector and aav vectors expressing reporter genes into mouse hippocampus. the adenovirus vector induced a pronounced immune response with a marked increase in cd45 and ... | 2004 | 14977565 |
| reengineered salivary glands are stable endogenous bioreactors for systemic gene therapeutics. | the use of critical-for-life organs (e.g., liver or lung) for systemic gene therapeutics can lead to serious safety concerns. to circumvent such issues, we have considered salivary glands (sgs) as an alternative gene therapeutics target tissue. given the high secretory abilities of sgs, we hypothesized that administration of low doses of recombinant adeno-associated virus (aav) vectors would allow for therapeutic levels of transgene-encoded secretory proteins in the bloodstream. we administered ... | 2004 | 14978265 |
| recombinant aav-mediated gene delivery to the central nervous system. | various regions of the brain have been successfully transduced by recombinant adeno-associated virus (raav) vectors with no detected toxicity. when using the cytomegalovirus immediate early (cmv) promoter, a gradual decline in the number of transduced cells has been described. in contrast, the use of cellular promoters such as the neuron-specific enolase promoter or hybrid promoters such as the chicken beta-actin/cmv promoter resulted in sustained transgene expression. the cellular tropism of ra ... | 2004 | 14978764 |
| transfection-free and scalable recombinant aav vector production using hsv/aav hybrids. | adeno-associated virus (aav) vectors are highly efficient tools for use in gene therapy. current production methods rely on plasmid transfection and are not generally considered amenable to scale-up. to improve recombinant aav (raav) vector production in terms of both final titre and simplicity, we constructed recombinant herpes simplex virus (hsv) vectors, either disabled (icp27 deleted) or nondisabled, encoding the aav rep and cap genes. we also integrated an raavgfp construct into the nondisa ... | 2004 | 14985784 |
| [adeno-associated virus mediated delivery and expression of gdnf cdna in cultured bovine iris pigment epithelial cells]. | to obtain transgenic bovine iris pigment epithelial cells (ipecs) by adeno-associated virus (aav) mediated delivery of cdna of glial cell-line derived neurotrophic factor (gdnf). | 2004 | 14989961 |
| distinct classes of proteasome-modulating agents cooperatively augment recombinant adeno-associated virus type 2 and type 5-mediated transduction from the apical surfaces of human airway epithelia. | tripeptidyl aldehyde proteasome inhibitors have been shown to effectively increase viral capsid ubiquitination and transduction of recombinant adeno-associated virus type 2 (raav-2) and raav-5 serotypes. in the present study we have characterized a second class of proteasome-modulating agents (anthracycline derivatives) for their ability to induce raav transduction. the anthracycline derivatives doxorubicin and aclarubicin were chosen for analysis because they have been shown to interact with th ... | 2004 | 14990705 |
| rapid uncoating of vector genomes is the key to efficient liver transduction with pseudotyped adeno-associated virus vectors. | transduction of the liver with single-stranded adeno-associated virus serotype 2 (aav2) vectors is inefficient; less than 10% of hepatocytes are permissive for stable transduction, and transgene expression is characterized by a lag phase of up to 6 weeks. aav2-based vector genomes packaged inside aav6 or aav8 capsids can transduce the liver with higher efficiency, but the molecular mechanisms underlying this phenomenon have not been determined. we now show that the primary barrier to transductio ... | 2004 | 14990730 |
| chronic phospholamban inhibition prevents progressive cardiac dysfunction and pathological remodeling after infarction in rats. | ablation or inhibition of phospholamban (pln) has favorable effects in several genetic murine dilated cardiomyopathies, and we showed previously that a pseudophosphorylated form of pln mutant (s16epln) successfully prevented progressive heart failure in cardiomyopathic hamsters. in this study, the effects of pln inhibition were examined in rats with heart failure after myocardial infarction (mi), a model of acquired disease. s16epln was delivered into failing hearts 5 weeks after mi by transcoro ... | 2004 | 14991071 |
| c-flip-l recombinant adeno-associated virus vector infection prevents fas-mediated but not nerve growth factor withdrawal-mediated cell death in pc12 cells. | fas is a cell surface death receptor that may play an important role in regulating cell death in neuronal cell types by activation of caspase 8. cellular flice inhibitory protein-long (c-flip-l) is an endogenous inhibitor of the activation of caspase 8 by fas. the current study addresses the role of c-flip-l in regulation of cell death in pc12 cells induced by nerve growth factor (ngf) withdrawal and fas antibody, which acts as a fas ligand and activates the fas receptor. a recombinant adeno-ass ... | 2004 | 14992818 |
| a parvovirus isolated from royal python (python regius) is a member of the genus dependovirus. | parvoviruses were isolated from python regius and boa constrictor snakes and propagated in viper heart (vh-2) and iguana heart (igh-2) cells. the full-length genome of a snake parvovirus was cloned and both strands were sequenced. the organization of the 4432-nt-long genome was found to be typical of parvoviruses. this genome was flanked by inverted terminal repeats (itrs) of 154 nt, containing 122 nt terminal hairpins and contained two large open reading frames, encoding the non-structural and ... | 2004 | 14993638 |
| prospects for gene therapy in the fragile x syndrome. | "if politics is the art of the possible, research is the art of the soluble. both are immensely practical-minded affairs." p. b. medawar.gene therapy is unarguably the definitive way to treat, and possibly cure, genetic diseases. a straightforward concept in theory, in practice it has proven difficult to realize, even when directed to easily accessed somatic cell systems. gene therapy for diseases in which the central nervous system (cns) is the target organ presents even greater challenges and ... | 2004 | 14994292 |
| adeno-associated virus 2-mediated antiangiogenic cancer gene therapy: long-term efficacy of a vector encoding angiostatin and endostatin over vectors encoding a single factor. | angiogenesis is characteristic of solid tumor growth and a surrogate marker for metastasis in many human cancers. inhibition of tumor angiogenesis using antiangiogenic drugs and gene transfer approaches has suggested the potential of this form of therapy in controlling tumor growth. however, for long-term tumor-free survival by antiangiogenic therapy, the factors controlling tumor neovasculature need to be systemically maintained at stable therapeutic levels. here we show sustained expression of ... | 2004 | 14996740 |
| adeno-associated virus rep78/rep68 promotes localized melting of the rep binding element in the absence of adenosine triphosphate. | we have applied fluorescence anisotropy and molecular beacon fluorescence methods to study the interactions between the adeno-associated virus rep78/rep68 protein and the 23-bp rep binding element (rbe). rep78/rep68 stably interacted with both the single- and double-stranded conformations of the rbe, but the interaction mechanisms of single- and double-stranded dna appeared to be fundamentally different. the stoichiometry of rep78 association with both the separate top and bottom strands of the ... | 2004 | 14997524 |
| aav-encoded expression of trail in experimental human colorectal cancer leads to tumor regression. | gene transfer vectors based on the adeno-associated virus (aav) are used for various experimental and clinical therapeutic approaches. in the present study, we demonstrate the utility of raav as a tumoricidal agent in human colorectal cancer. we constructed an raav vector that expresses tumor necrosis factor (tnf)-related apoptosis-inducing ligand (trail/apo2l) and used it to transduce human colorectal cancer cells. trail belongs to the tnf superfamily of cytokines that are involved in various i ... | 2004 | 14999225 |
| rep68 protein of adeno-associated virus type 2 interacts with 14-3-3 proteins depending on phosphorylation at serine 535. | rep78/68 proteins of adeno-associated virus type 2 (aav-2) are involved in many aspects of the viral life cycle, including replication, gene expression, and site-specific integration. to understand the molecular mechanisms of the actions of rep proteins, we searched for rep68-interacting cellular proteins by utilizing a one-step affinity purification technique and identified two members of 14-3-3 proteins (14-3-3 epsilon and gamma). we found that phosphorylation of 535ser at the carboxy terminus ... | 2004 | 15003870 |
| human papillomavirus, cytomegalovirus, and adeno-associated virus infections in pregnant and nonpregnant women with cervical intraepithelial neoplasia. | two hundred eight cervical specimens from two groups of subjects, 165 nonpregnant women and 53 pregnant women with cervical intraepithelial neoplasia (cin) of grades i to iii, were positive by pcr analyses for human papillomaviruses (hpvs), adeno-associated virus type 2 (aav 2), and human cytomegalovirus (hcmv) in 67, 6, and 4.1% of the cases, respectively. the presence of aav 2 infection was more frequently associated with pregnancy (17 versus 2.4%) and hpv-positive cervices (odds ratio = 6.358 ... | 2004 | 15004114 |
| efficient delivery of cre-recombinase to neurons in vivo and stable transduction of neurons using adeno-associated and lentiviral vectors. | inactivating genes in vivo is an important technique for establishing their function in the adult nervous system. unfortunately, conventional knockout mice may suffer from several limitations including embryonic or perinatal lethality and the compensatory regulation of other genes. one approach to producing conditional activation or inactivation of genes involves the use of cre recombinase to remove loxp-flanked segments of dna. we have studied the effects of delivering cre to the hippocampus an ... | 2004 | 15005815 |
| role of viral vectors and virion shells in cellular gene expression. | the role of the virion shell in viral pathogenesis is relatively unknown yet the use of viral vectors in human gene transfer experiments requires an understanding of these interactions. in this study, we used dna microarrays to identify genes modulated during pathogenic adenovirus or nonpathogenic adeno-associated virus infections. responses to wt viruses, recombinant vectors, or empty virion particles were compared. adeno-associated virus shells induced nearly the full complement of changes eli ... | 2004 | 15006600 |
| efficient gene transfer of hiv-1-specific short hairpin rna into human lymphocytic cells using recombinant adeno-associated virus vectors. | the cellular introduction of short, interfering rna leads to sequence-specific degradation of homologous mrna, a process termed rna interference (rnai). here, we report that recombinant adeno-associated virus 2 (raav-2) can be used to transfer short hairpin (sh) rna expression cassettes genetically into human cells. hiv-1 replication was suppressed by >95% in h9 cells and primary human lymphocytes that expressed shrna targeting the first exon of the viral transactivator protein tat compared to c ... | 2004 | 15006606 |
| optimal design of a single recombinant adeno-associated virus derived from serotypes 1 and 2 to achieve more tightly regulated transgene expression from nonhuman primate muscle. | recombinant adeno-associated virus (raav) vector supports long-term transgene expression from skeletal muscle in most mammals, including human. in some instances, the requirement for tight control of the transgene expression is expected. the original tetracycline-dependent system using the rtta (dox-on) transactivator displayed a baseline activity in the off state but improved versions are now available and need to be evaluated in a single-raav-vector strategy. in the present study we cloned, in ... | 2004 | 15006608 |
| a new method for recombinant adeno-associated virus vector delivery to murine diaphragm. | genetically modified mice are important models for evaluation of potential gene therapies for human diseases. however, their small size often precludes the use of clinically feasible methods for vector delivery, therefore, alternative methods must be used. we have developed a gel-based method for delivery of recombinant adeno-associated virus vectors to the mouse diaphragm, an important target organ for many myopathic diseases. we hypothesized that delivery of vectors in a viscous solution would ... | 2004 | 15006614 |
| structure of adeno-associated virus serotype 5. | adeno-associated virus serotype 5 (aav5) requires sialic acid on host cells to bind and infect. other parvoviruses, including aleutian mink disease parvovirus (adv), canine parvovirus (cpv), minute virus of mice, and bovine parvovirus, also bind sialic acid. hence, structural homology may explain this functional homology. the amino acids required for cpv sialic acid binding map to a site at the icosahedral twofold axes of the capsid. in contrast to aav5, aav2 does not bind sialic acid, but rathe ... | 2004 | 15016858 |
| direct exposure of mouse spermatozoa to very high concentrations of a serotype-2 adeno-associated virus gene therapy vector fails to lead to germ cell transduction. | in a clinical safety trial involving an adeno-associated virus (aav) gene therapy vector encoding human factor ix, intrahepatic administration of the vector was associated with the finding of vector dna in semen that persisted for several weeks. uncertainty remains as to the route by which the vector reached semen, but the finding raised the prospect that mature sperm could be exposed to the vector and sustain integration of vector dna. to provocatively test for the ability of aav vectors to tra ... | 2004 | 15018737 |
| immune response against gene therapy vectors: influence of synovial fluid on adeno-associated virus mediated gene transfer to chondrocytes. | intraarticular gene transfer with adeno-associated virus (aav) vectors may allow efficient therapeutic transgene expression within the joint. in an effort to understand potential obstacles (particularly immunity against aav vectors) to intraarticular gene therapy better, our objective was to determine whether synovial fluid (sf) influenced aav-mediated gene transfer to chondrocytes. sf and sera from 21 patients with joint diseases were collected. neutralizing activity against aav/interleukin-4 ( ... | 2004 | 15024183 |
| pre-emptive gene therapy using recombinant adeno-associated virus delivery of extracellular superoxide dismutase protects heart against ischemic reperfusion injury, improves ventricular function and prolongs survival. | in high-risk patients, the ideal cardiovascular gene therapy requires a strategy that provides long-term protection of myocardium against episodes of ischemic/reperfusion injury. we report the development of an efficient, long-lasting pre-emptive gene therapy strategy in a rat model of ischemic-reperfusion (i/r) injury of heart. at 6 weeks prior to myocardial injury, the human extracellular superoxide dismutase (ec-sod) gene was delivered by direct intramyocardial injections, using a recombinant ... | 2004 | 15029230 |
| [gene therapy for muscular dystrophy]. | duchenne muscular dystrophy (dmd) is an x-linked, lethal muscle disorder caused by mutations in the dystrophin gene. although an adeno-associated virus (aav) vector-mediated gene transfer provides an attractive approach to the treatment of dmd, limitation in insertion size up to 4.9 kb excludes incorporation of a full-length dystrophin cdna (14 kb) into an aav vector. we previously generated micro-dystrophin transgenic dystrophin-deficient mdx mice. in 4.9 kb rod-truncated micro-dystrophin cs1 t ... | 2004 | 15031985 |
| alpha-fodrin is cleaved by caspase-3 in a chronic ocular hypertensive (coh) rat model of glaucoma. | alpha-fodrin is a neuronal cytoskeletal protein and a known caspase-3 target. we sought to determine whether caspase-3 cleaves alpha-fodrin in coh rat retinas and whether this process is reduced by adeno-associated virus (aav)-induced retinal ganglion cell expression of baculovirus inhibitory repeat-containing 4 (birc4), a potent caspase-3 inhibitor. | 2004 | 15036563 |
| possible role of bacterial and viral infections in miscarriages. | to determine the role of infections in miscarriages. chorionic villi from aborted material were subjected to cytogenetic evaluation and analyzed for the presence of chlamydia trachomatis, ureaplasma urealyticum, mycoplasma hominis, human cytomegalovirus (hcmv), adeno-associated virus (aav), and human papillomaviruses (hpv). | 2004 | 15037417 |
| gene therapy progress and prospects: recombinant adeno-associated virus (raav) vectors. | 2004 | 15042119 | |
| anticonvulsant and antiepileptogenic effects mediated by adeno-associated virus vector neuropeptide y expression in the rat hippocampus. | neuropeptide y (npy) inhibits seizures in experimental models and reduces excitability in human epileptic tissue. we studied the effect of long-lasting npy overexpression in the rat hippocampus with local application of recombinant adeno-associated viral (aav) vectors on acute kainate seizures and kindling epileptogenesis. transgene expression was significantly increased by 7 d, reached maximal expression by 2 weeks, and persisted for at least 3 months. serotype 2 aav vector increased npy expres ... | 2004 | 15044544 |
| targeted correction of single-base-pair mutations with adeno-associated virus vectors under nonselective conditions. | recombinant adeno-associated virus (raav) vectors possess the unique ability to introduce genetic alterations at sites of homology in genomic dna through a mechanism thought to predominantly involve homologous recombination. we have investigated the efficiency of this approach using a mutant enhanced green fluorescent protein (egfp) fluorescence recovery assay that facilitates detection of gene correction events in living cells under nonselective conditions. our data demonstrate that raav infect ... | 2004 | 15047832 |
| gene therapy with human recombinant osteoprotegerin reverses established osteopenia in ovariectomized mice. | osteoporosis is a chronic condition that is typically treated by the long-term repeated administration of antiresorptive agents. gene therapy has the potential to deliver protein-based antiresorptive agents without the need for repeated administration. osteoprotegerin (opg) is a naturally occuring protein that prevents bone resorption by inhibiting osteoclast formation, function and survival. we tested whether adeno-associated virus (aav) could deliver opg at levels that are sufficient to revers ... | 2004 | 15050896 |
| stable transduction of large dna by high-capacity adeno-associated virus/adenovirus hybrid vectors. | viral vectors with high cloning capacity and host chromosomal integration ability are in demand for the efficient and permanent genetic modification of target cells with large dna molecules. we have generated a hybrid gene transfer vehicle consisting of recombinant adeno-associated virus (aav) replicative intermediates packaged in adenovirus (ad) capsids. this arrangement allows cell cycle-independent nuclear delivery of recombinant aav genomes with lengths considerably above the maximum size (i ... | 2004 | 15051388 |
| comparison of adenoviral and adeno-associated viral vectors for pancreatic gene delivery in vivo. | although effective gene therapy vectors have been developed for organ systems such as the liver, an effective delivery vector to the pancreas in vivo has remained elusive. of the currently available viral vectors, adenovirus and adeno-associated virus (aav) are two of the most efficient at transducing nondividing cells. we have constructed recombinant adenovirus (advlacz), adeno-associated virus serotype 2 (aav2lacz), and pseudotyped adeno-associated virus serotype 5 and 8 (aav5lacz, aav8lacz) c ... | 2004 | 15053865 |
| long-term correction of hyperphenylalaninemia by aav-mediated gene transfer leads to behavioral recovery in phenylketonuria mice. | classical phenylketonuria (pku) is a metabolic disorder caused by a deficiency of the hepatic enzyme phenylalanine hydroxylase (pah). if untreated, accumulation of phenylalanine will damage the developing brain of affected individuals, leading to severe mental retardation. here, we show that a liver-directed pah gene transfer brought about long-term correction of hyperphenylalaninemia and behavioral improvement in a mouse model of pku. a recombinant adeno-associated virus (aav) vector carrying t ... | 2004 | 15057263 |
| novel therapeutic approach for hemophilia using gene delivery of an engineered secreted activated factor vii. | hemophilia is a bleeding disorder caused by mutations in the genes encoding coagulation factor viii (fviii) or fix. current treatment is through intravenous infusion of the missing protein. the major complication of treatment is the development of neutralizing ab's to the clotting factor. infusion of recombinant activated human factor vii (rhfviia), driving procoagulant reactions independently of human fviii (hfviii) or hfix, has been successful in such patients and could in theory provide hemos ... | 2004 | 15057309 |
| recombinant adeno-associated virus as delivery vector for gene therapy--a review. | recombinant adeno-associated virus (raav) is one of the most promising delivery vectors for gene therapy, due to its nonpathogenic property, nonimmunogenecity to host, and broad cell and tissue tropisms. this article summarizes the biological characteristics of aav; the procedures to prepare, purify, and characterize the raav for gene therapy applications; and some of the clinical trials utilizing raav as delivery vehicles. also discussed are the current efforts to modify raav to change its trop ... | 2004 | 15068701 |
| adeno-associated virus mediated endostatin gene therapy in combination with topoisomerase inhibitor effectively controls liver tumor in mouse model. | raav mediated endostatin gene therapy has been examined as a new method for treating cancer. however, a sustained and high protein delivery is required to achieve the desired therapeutic effects. we evaluated the impact of topoisomerase inhibitors in raav delivered endostatin gene therapy in a liver tumor model. | 2004 | 15069724 |
| cross-dressing the virion: the transcapsidation of adeno-associated virus serotypes functionally defines subgroups. | for all adeno-associated virus (aav) serotypes, 60 monomers of the vp1, vp2, and vp3 structural proteins assemble via an unknown mechanism to form an intact capsid. in an effort to better understand the properties of the capsid monomers and their role in viral entry and infection, we evaluated whether monomers from distinct serotypes can be mixed to form infectious particles with unique phenotypes. this transcapsidation approach consisted of the transfection of pairwise combinations of aav serot ... | 2004 | 15078923 |
| total correction of hemophilia a mice with canine fviii using an aav 8 serotype. | despite the popularity of adeno-associated virus 2 (aav2) as a vehicle for gene transfer, its efficacy for liver-directed gene therapy in hemophilia a or b has been suboptimal. here we evaluated aav serotypes 2, 5, 7, and 8 in gene therapy of factor viii (fviii) deficiency in a hemophilia a mouse model and found that aav8 was superior to the other 3 serotypes. we expressed canine b domain-deleted fviii cdna either in a single vector or in 2 separate aav vectors containing the heavy- and light-ch ... | 2004 | 14551134 |
| impurity of recombinant adeno-associated virus type 2 affects the transduction characteristics following subretinal injection in the rat. | we recently reported that different purification methods of recombinant adeno-associated virus type 2 (raav2) affect the transduction characteristics following subretinal injection. in this study, we examined the roles of contaminant proteins from the hek-293 cells and helper adenovirus, inactivation of helper adenovirus and cell stress induced by dna-damaging agents in raav-mediated retinal transduction. our results showed that contaminating factors/proteins resulting from the helper e1 deleted ... | 2004 | 14659960 |
| alternative polyadenylation of adeno-associated virus type 5 rna within an internal intron is governed by both a downstream element within the intron 3' splice acceptor and an element upstream of the p41 initiation site. | adeno-associated virus type 5 (aav5) has a linear, single-stranded dna genome of ca. 5 kb and an overlapping transcription profile featuring multiple promoters and a single intron in the center of the genome. unlike the situation for the prototype aav2, aav5 rnas transcribed from upstream promoters at map units 7 (p7) and 19 (p19), which encode the viral rep proteins, are predominantly polyadenylated at a site within the intron [(pa)p]. rnas generated from the aav5 capsid gene promoter p41, whic ... | 2004 | 14671090 |
| spatial and temporal organization of adeno-associated virus dna replication in live cells. | upon cell entry, the genomes of herpes simplex virus type 1 (hsv-1) and adenovirus (ad) associate with distinct nuclear structures termed nd10 or promyelocytic leukemia (pml) nuclear bodies (nbs). pml nb morphology is altered or disrupted by specific viral proteins as replication proceeds. we examined whether adeno-associated virus (aav) replication compartments also associate with pml nbs, and whether modification or disruption of these by hsv-1 or ad, both of which are helper viruses for aav, ... | 2004 | 14671120 |
| the rep protein of adeno-associated virus type 2 interacts with single-stranded dna-binding proteins that enhance viral replication. | adeno-associated virus (aav) type 2 is a human parvovirus whose replication is dependent upon cellular proteins as well as functions supplied by helper viruses. the minimal herpes simplex virus type 1 (hsv-1) proteins that support aav replication in cell culture are the helicase-primase complex of ul5, ul8, and ul52, together with the ul29 gene product icp8. we show that aav and hsv-1 replication proteins colocalize at discrete intranuclear sites. transfections with mutant genes demonstrate that ... | 2004 | 14671124 |
| heat-shock treatment-mediated increase in transduction by recombinant adeno-associated virus 2 vectors is independent of the cellular heat-shock protein 90. | recombinant adeno-associated virus 2 (aav) vectors transduction efficiency varies greatly in different cell types. we have described that a cellular protein, fkbp52, in its phosphorylated form interacts with the d-sequence in the viral inverted terminal repeat, inhibits viral second strand dna synthesis, and limits transgene expression. here we investigated the role of cellular heat-shock protein 90 (hsp90) in aav transduction because fkbp52 forms a complex with hsp90, and because heat-shock tre ... | 2004 | 14711833 |
| recombinant adeno-associated virus-mediated alpha-1 antitrypsin gene therapy prevents type i diabetes in nod mice. | type i diabetes results from an autoimmune destruction of the insulin-producing pancreatic beta cells. although the exact immunologic processes underlying this disease are unclear, increasing evidence suggests that immunosuppressive, immunoregulatory and anti-inflammatory agents can interrupt the progression of the disease. alpha 1 antitrypsin (aat) is a multifunctional serine proteinase inhibitor (serpin) that also displays a wide range of anti-inflammatory properties. to test the ability of aa ... | 2004 | 14712302 |
| intravenous administration of an aav-2 vector for the expression of factor ix in mice and a dog model of hemophilia b. | previous experiments have demonstrated the stable expression of factor ix (fix) protein in mice and canine models of hemophilia b following portal vein gene transfer with a recombinant adeno-associated virus (raav) vector encoding fix. here, we present the results of studies that further optimized the raav vector transgene cassette used to express fix and explored the use of the less-invasive intravenous (i.v.) route of vector administration for the treatment of hemophilia b. first, a liver-spec ... | 2004 | 14712305 |
| an overview of current delivery systems in cancer gene therapy. | the main objective in gene therapy is the development of efficient, non-toxic gene carriers that can encapsulate and deliver foreign genetic materials into specific cell types such as cancerous cells. during the past two decades, enormous research in the area of gene delivery has been conducted worldwide, in particular for cancer gene therapy application. viral vectors are biological systems derived from naturally evolved viruses capable of transferring their genetic materials into the host cell ... | 2004 | 14684267 |
| can genes transduced by adeno-associated virus vectors elicit or evade an immune response? | vectors derived from the adeno-associated viruses (aav) have been successfully used for the long-term expression of therapeutic genes in animal models and in patients. stable expression of the transgene in the transduced cells can be used to correct disease stemming from genetic deficiencies. one of the major advantages of these vectors is reported to be the absence of deleterious immune responses following gene transfer. however, recent studies have shown that aav vectors elicit humoral and cel ... | 2004 | 14689272 |
| erythropoietin gene therapy leads to autoimmune anemia in macaques. | gene therapy is being considered for the delivery of therapeutic proteins. we evaluated the delivery of the hormone erythropoietin (epo) into cynomolgus macaques through intramuscularly administered adeno-associated virus (aav) vectors. as expected, the animals developed supraphysiologic levels of epo and polycythemia. however, severe anemia ensued in some animals because of an autoimmune response to endogenous and transgene derived epo. this is the first example of gene therapy leading to inadv ... | 2004 | 14695227 |
| potential for germ line transmission after intramyocardial gene delivery by adeno-associated virus. | intramyocardial injection of adeno-associated virus (aav) has been shown to be an effective strategy for cardiac gene delivery. this approach leads to long-term gene expression in the heart, offering the possibility of chronic gene therapy. however, the long-term safety of this approach with regard to vector bio-distribution and extracardiac transgene expression has not been evaluated. to examine these issues, 8-week-old male sprague-dawley rats were injected intramyocardially with either 4x10(1 ... | 2004 | 14697221 |
| viral vector-mediated gene transfer of neurotrophins to promote regeneration of the injured spinal cord. | injuries to the adult mammalian spinal cord often lead to severe damage to both ascending (sensory) pathways and descending (motor) nerve pathways without the perspective of complete functional recovery. future spinal cord repair strategies should comprise a multi-factorial approach addressing several issues, including optimalization of survival and function of spared central nervous system neurons in partial lesions and the modulation of trophic and inhibitory influences to promote and guide ax ... | 2004 | 14699980 |
| parvovirus vectors for cancer gene therapy. | parvoviruses comprise a group of single-stranded dna viruses with greater potential for gene therapy applications. unique characteristics of paroviruses, such as non-pathogenicity, antioncogenicity and methods of efficient recombinant vector production, have drawn more attention towards utilising parvovirus-based vectors in cancer gene therapy. although > 30 different parvoviruses have been identified so far, recombinant vectors derived from adeno-associated virus (aav), minute virus of mice (mv ... | 2004 | 14680468 |
| light-activated gene transduction of recombinant adeno-associated virus in human mesenchymal stem cells. | deficiencies in skeletal tissue repair and regeneration lead to conditions like osteoarthritis, osteoporosis and degenerative disc disease. while no cure for these conditions is available, the use of human bone marrow derived-mesenchymal stem cells (humscs) has been shown to have potential for cell-based therapy. furthermore, recombinant adeno-associated viruses (raav) could be used together with humscs for in vivo or ex vivo gene therapy. unfortunately, the poor transduction efficiency of these ... | 2004 | 14681695 |
| tumor-specific gene expression using regulatory elements of the glucose transporter isoform 1 gene. | in order to achieve tumor-specific targeting of adeno-associated virus (aav)-mediated gene expression, the promoter of the glucose transporter isoform 1 (glut1) gene was cloned upstream of the enhanced green fluorescence protein (egfp) and the herpes simplex virus thymidine kinase (hsvtk) gene. facs analysis performed at 48 h after transient infection with raav/cytomegalovirus (cmv)egfp viral particles revealed an increase of fluorescence in all the cell lines tested. however, egfp expression un ... | 2004 | 14681725 |
| aav-mediated gene transfer of tissue inhibitor of metalloproteinases-1 inhibits vascular tumor growth and angiogenesis in vivo. | the activity of matrix metalloproteinases (mmps) is a universal feature of cellular invasion, tumor angiogenesis and metastasis, which is counterbalanced and regulated by the natural tissue inhibitors of mmps (timps). here we show that timp1 gene transfer delivered by an adeno-associated virus (aav) vector inhibits tumor growth in a murine xenotransplant model. a human kaposi's sarcoma cell line, forming highly vascularized tumors in vivo and having a high natural permissivity to aav gene transf ... | 2004 | 14681728 |
| development of packaging cell lines for generation of adeno-associated virus vectors by lentiviral gene transfer of trans-complementary components. | adeno-associated virus (aav) vector system has several useful advantages with regard to in vitro and in vivo gene transfer. however, their usages have been limited by cumbersome and labor-intensive vector production in the traditional method. to overcome limitations in aav production, in this report, we explored the possibility of generating aav packaging cell line, 293t r/c.va.e2a.e4. cells, by using lentivirus-mediated transduction of rep/cap gene of aav-2, va rna, e2a, and e4 genes of ad5 int ... | 2004 | 15347316 |
| [immortalization of human embryonic cervical epithelial cells induced by e6, e7 genes of human papillomavirus 16]. | to establish an immortalized cell line derived from the embryonic cervical epithelium by infection with the recombinant adeno-associated virus (raav) containing human papillomavirus (hpv)16 e6, e7, and to study the biological features of cervical cancer cell line. | 2004 | 15347475 |
| characterization of a nuclear localization signal in the c-terminus of the adeno-associated virus rep68/78 proteins. | adeno-associated virus (aav) replicates in the nucleus of infected cells, and therefore multiple nuclear import events are required for productive infection. we analyzed nuclear import of the viral rep proteins and characterized a nuclear localization signal (nls) in the c-terminus. we demonstrate that basic residues in this region constitute an nls that is transferable and mediates interaction with the nuclear import receptor importin alpha in vitro. mutant rep proteins are predominantly cytopl ... | 2004 | 15351208 |
| sequence analysis, viral rescue from infectious clones and generation of recombinant virions of the avian adeno-associated virus. | aiming at the generation of a viral-vectored system for gene delivery and vaccination in poultry, the entire genomes of the vr-865 and da-1 strains of the avian adeno-associated virus have been cloned and sequenced. sequence analysis of the clones showed that the genomic distribution of the structural and non-structural protein-coding genes of these viruses is conserved and in agreement with what has been previously described for the primate adeno-associated viruses. amino acid differences betwe ... | 2004 | 15351493 |
| differential raav2 transduction efficiencies and insulin secretion profiles in pure and co-culture models of human enteroendocrine l-cells and enterocytes. | cell-based therapies for treating insulin-dependent diabetes (idd) can provide a more physiologic regulation of blood glucose levels in a less invasive fashion than insulin injections. previously, we developed an engineered human enteroendocrine l-cell model for regulated insulin release via recombinant adeno-associated virus serotype 2, or raav2, transduction. the aim of this study was to evaluate the efficiency and selectivity of raav2-mediated insulin gene delivery to enteroendocrine l-cells ... | 2004 | 15352073 |