| titration of aav-2 particles via a novel capsid elisa: packaging of genomes can limit production of recombinant aav-2. | we demonstrate the rapid and reliable quantification of physical aav-2 (adeno-associated virus type 2) particles via a novel elisa based on a monoclonal antibody which selectively recognizes assembled aav-2 capsids. titration of a variety of recombinant aav-2 (raav) preparations revealed that at least 80+percent of all particles were empty, compared with a maximum of 50percent in wild-type aav-2 stocks, indicating that the recombinant genomes were less efficiently encapsidated. this finding was ... | 1999 | 10455443 |
| cellular redox state alters recombinant adeno-associated virus transduction through tyrosine phosphatase pathways. | several types of environmental damage including uv, hydroxyurea and ionizing irradiation have been shown to augment raav transduction. current hypotheses suggest that these environmental stimuli lead to the enhanced production and/or activation of cellular factors important in the conversion of single-stranded dna genomes to expressible forms. however, the mechanisms of action are currently unknown. we hypothesized that reactive oxygen intermediates (roi) may play a common role in the augmentati ... | 1999 | 10467367 |
| evidence for infection of the human embryo with adeno-associated virus in pregnancy. | previous reports have demonstrated the presence of dna of the human helper virus-dependent adeno-associated parvovirus (aav) in uterine tissue and curettage material from early miscarriage. to examine infection of embryonic tissue during pregnancy, amnion fluids were analysed for the presence of aav. using polymerase chain reaction, aav dna was detected in 64 out of 238 dna samples extracted from amnion cells. dna of helper viruses were found in 12% (papillomavirus) and 18% (cytomegalovirus) of ... | 1999 | 10469719 |
| genetic capsid modifications allow efficient re-targeting of adeno-associated virus type 2. | the human parvovirus adeno-associated virus type 2 (aav2) has many features that make it attractive as a vector for gene therapy. however, the broad host range of aav2 might represent a limitation for some applications in vivo, because recombinant aav vector (raav)-mediated gene transfer would not be specific for the tissue of interest. this host range is determined by the binding of the aav2 capsid to specific cellular receptors and/or co-receptors. the tropism of aav2 might be changed by genet ... | 1999 | 10470084 |
| transduction of renal cells in vitro and in vivo by adeno-associated virus gene therapy vectors. | there has been an increasing interest recently in the possibility of treating renal diseases using gene therapy. the ability to pursue gene therapy for renal diseases has been limited by the availability of an adequate system for gene delivery to the kidney. adeno-associated virus (aav) is a defective virus of the parvovirus family that has a number of properties attractive for renal gene delivery: recombinant aav contains no viral genes; expression of genes delivered by these vectors does not a ... | 1999 | 10477142 |
| human herpesvirus 6b genome sequence: coding content and comparison with human herpesvirus 6a. | human herpesvirus 6 variants a and b (hhv-6a and hhv-6b) are closely related viruses that can be readily distinguished by comparison of restriction endonuclease profiles and nucleotide sequences. the viruses are similar with respect to genomic and genetic organization, and their genomes cross-hybridize extensively, but they differ in biological and epidemiologic features. differences include infectivity of t-cell lines, patterns of reactivity with monoclonal antibodies, and disease associations. ... | 1999 | 10482553 |
| factors affecting the terminal resolution site endonuclease, helicase, and atpase activities of adeno-associated virus type 2 rep proteins. | the rep68 and rep78 proteins (rep68/78) of adeno-associated virus type 2 (aav) are critical for aav replication and site-specific integration. they bind specifically to the aav inverted terminal repeats (itrs) and possess atpase, helicase, and strand-specific/site-specific endonuclease activities. in the present study, we further characterized the aav rep68/78 helicase, atpase, and endonuclease activities by using a maltose binding protein-rep68 fusion (mbp-rep68delta) produced in escherichia co ... | 1999 | 10482574 |
| latent adeno-associated virus infection elicits humoral but not cell-mediated immune responses in a nonhuman primate model. | latent infection with wild-type (wt) adeno-associated virus (aav) was studied in rhesus macaques, a species that is a natural host for aav and that has some homology to humans with respect to the preferred locus for wt aav integration. each of eight animals was infected with an inoculum of 10(10) iu of wt aav, administered by either the intranasal, intramuscular, or intravenous route. two additional animals were infected intranasally with wt aav and a helper adenovirus (ad), while one additional ... | 1999 | 10482608 |
| structural analysis of adeno-associated virus transduction circular intermediates. | recombinant adeno-associated virus (raav) has recently been demonstrated to form circular intermediates following transduction in muscle tissue and cell lines. although restriction enzyme and southern blot analysis has revealed a consistent monomer and multimer head-to-tail conformation, detailed structural sequence analysis has been lacking due to the high secondary structure of the itr arrays. to gain further insight into potential mechanisms by which aav circular genomes are formed from linea ... | 1999 | 10484751 |
| progress in direct striatal delivery of l-dopa via gene therapy for treatment of parkinson's disease using recombinant adeno-associated viral vectors. | viral vectors have recently been used successfully to transfer genes and express different proteins in the brain. this review discusses the requirements to consider human clinical trials in which recombinant adeno-associated virus vectors are used to transfer the genes necessary to produce l-dihydroxyphenylalanine (l-dopa) directly into the striatum of parkinson's patients. preclinical data that apply to the criteria defined as prerequisite for clinical trials are discussed. thus, in animal mode ... | 1999 | 10486174 |
| relative hypoglycemia and hyperinsulinemia in mice with heterozygous lipoprotein lipase (lpl) deficiency. islet lpl regulates insulin secretion. | lipoprotein lipase (lpl) provides tissues with fatty acids, which have complex effects on glucose utilization and insulin secretion. to determine if lpl has direct effects on glucose metabolism, we studied mice with heterozygous lpl deficiency (lpl+/-). lpl+/- mice had mean fasting glucose values that were up to 39 mg/dl lower than lpl+/+ littermates. despite having lower glucose levels, lpl+/- mice had fasting insulin levels that were twice those of +/+ mice. hyperinsulinemic clamp experiments ... | 1999 | 10488074 |
| immune responses to adenovirus and adeno-associated virus in humans. | vectors based on human adenovirus (ad) and adeno-associated virus (aav) are being evaluated for human gene therapy. the response of the host to the vector, in terms of antigen-specific immunity, will play a substantial role in clinical outcome. we have surveyed cohorts of normal subjects and cystic fibrosis patients for pre-existing immunity to these viruses, caused by naturally acquired infections. a number of humoral and cellular assays to adenovirus serotype 5 (ad5) and adeno-associated virus ... | 1999 | 10490767 |
| induction of apoptosis by cadmium and the adeno-associated virus rep proteins. | the parvoviruses exert antiproliferative effects on transformed cells in culture. the development of cell lines that inducibly express the parvovirus nonstructural proteins have implicated these proteins in the limitation of cell growth. to study the host cell interactions of the nonstructural proteins we have developed a human 293 cell line that expresses the adeno-associated virus (aav) rep gene upon induction with heavy metal salts. when induced with both zn(2+) and cd(2+), rep protein expres ... | 1999 | 10497113 |
| persistent, therapeutically relevant levels of human granulocyte colony-stimulating factor in mice after systemic delivery of adeno-associated virus vectors. | adeno-associated virus (aav) vectors have been shown to preferentially transduce hepatocytes after systemic administration in adult mice and to provide long-term expression of introduced genes. one application of this technology would be for the production of granulocyte colony-stimulating factor (g-csf), which increases mature neutrophil numbers in humans and in animals, and has therapeutic effects in disorders featuring chronic neutropenia, including cyclic, severe congenital, and idiopathic n ... | 1999 | 10498245 |
| enzymatic correction and cross-correction of mucopolysaccharidosis type i fibroblasts by adeno-associated virus-mediated transduction of the alpha-l-iduronidase gene. | mucopolysaccharidosis type i (mps i), a deficiency in the lysosomal enzyme alpha-l-iduronidase (idua), is characterized by skeletal abnormalities, hepatosplenomegaly and neurological dysfunction. to evaluate the potential for treatment of the disease using a gene delivery approach, recombinant adeno-associated virus (raav) vectors were constructed and evaluated for expression of the human idua cdna in transduced cells. 293 cells transduced with these aav vectors contained idua activity at 0.5 to ... | 1999 | 10498248 |
| in vivo marking of rhesus monkey lymphocytes by adeno-associated viral vectors: direct comparison with retroviral vectors. | we have compared adeno-associated virus (aav)-based and retrovirus-based vectors for their ability to transduce primary t lymphocytes in vitro and then tracked the persistence of these genetically marked lymphocytes in vivo, using the rhesus monkey model. to avoid the complication of immune rejection of lymphocytes transduced with xenogeneic genes in tracking studies primarily designed to investigate transduction efficiency and in vivo kinetics, the vectors were designed without expressed genes. ... | 1999 | 10498598 |
| submucosal gland development in the airway is controlled by lymphoid enhancer binding factor 1 (lef1). | previous studies have demonstrated that transcription of the lymphoid enhancer binding factor 1 (lef1) gene is upregulated in submucosal gland progenitor cells just prior to gland bud formation in the developing ferret trachea. in the current report, several animal models were utilized to functionally investigate the role of lef1 in initiating and supporting gland development in the airway. studies on lef1-deficient mice and antisense oligonucleotides in a ferret xenograft model demonstrate that ... | 1999 | 10498680 |
| gene transfer and models of gene therapy for the myocardium. | 1. gene transfer into the myocardium can be achieved through direct injection of plasmid dna or through the delivery of viral vectors, either directly or through the coronary vasculature. direct dna injection has proven extremely valuable in studies aimed at characterizing the activities of promoter elements in cardiac tissue and for examining the influence of the pathophysiological state of the myocardium on expression of transferred foreign genes. 2. viral vectors, in particular adenoviruses a ... | 1999 | 10499153 |
| highly regulated expression of adeno-associated virus large rep proteins in stable 293 cell lines using the cre/loxp switching system. | since the rep proteins of adeno-associated virus (aav) are harmful to cells, it is difficult to obtain stable cell lines that express them constitutively. in this study, stable 293 cell lines were obtained in which large rep expression was inducible by using the cre/loxp switching system. to determine the function of the induced rep proteins, the packaging capacity was examined after supplementation with a plasmid expressing small rep and cap proteins. a significant amount of recombinant aav (5. ... | 1999 | 10501504 |
| update on the prevalence of serum antibodies (igg and igm) to adeno-associated virus (aav). | in view of presumed non-pathogenicity, tumor suppressive properties, and site-specific integration of the viral genome the human parvovirus, adeno-associated virus (aav) has gained great interest as a gene transduction vector. data on the seroprevalence of antibodies to aav vary between reports, probably due to the different serological methods used. in order to understand better the immune response to aav during natural infection, sera from different age groups and various geographical regions ... | 1999 | 10502275 |
| enhanced expression of transgenes from adeno-associated virus vectors with the woodchuck hepatitis virus posttranscriptional regulatory element: implications for gene therapy. | the woodchuck hepatitis virus posttranscriptional regulatory element (wpre) evolved to stimulate the expression of intronless viral messages. to determine whether this ability to enhance expression could be useful in nonviral and heterologous viral gene delivery systems, we analyzed the ability of the wpre to elevate the expression of a cdna encoding the green fluorescent protein (gfp) in these contexts. we find that the wpre can stimulate the expression of gfp when the gene is delivered by tran ... | 1999 | 10515449 |
| adeno-associated virus type 2 protein interactions: formation of pre-encapsidation complexes. | the nonstructural adeno-associated virus type 2 rep proteins are known to control viral replication and thus provide the single-stranded dna genomes required for packaging into preformed capsids. in addition, complexes between rep proteins and capsids have previously been observed in the course of productive infections. such complexes have been interpreted as genome-linked rep molecules associated with the capsid upon successful dna encapsidation. here we demonstrate via coimmunoprecipitation, c ... | 1999 | 10516005 |
| integrating adenovirus-adeno-associated virus hybrid vectors devoid of all viral genes. | recently, we demonstrated that inverted repeat sequences inserted into first-generation adenovirus (ad) vector genomes mediate precise genomic rearrangements resulting in vector genomes devoid of all viral genes that are efficiently packaged into functional ad capsids. as a specific application of this finding, we generated adenovirus-adeno-associated virus (aav) hybrid vectors, first-generation ad vectors containing aav inverted terminal repeat sequences (itrs) flanking a reporter gene cassette ... | 1999 | 10516040 |
| rep-mediated nicking of the adeno-associated virus origin requires two biochemical activities, dna helicase activity and transesterification. | the single-stranded adeno-associated virus (aav) genome is flanked by terminal hairpinned origins of dna replication (terminal repeats [trs]) that are nicked at the terminal resolution site (trs) by the aav rep protein in an atp-dependent, site-specific manner. here we determine the minimal trs sequence necessary for rep cleavage, 3'-ccggt/tg-5', and show that this 7-base core sequence is required only on the nicked strand. we also identify a potential stem-loop structure at the trs. interesting ... | 1999 | 10516041 |
| charge-to-alanine mutagenesis of the adeno-associated virus type 2 rep78/68 proteins yields temperature-sensitive and magnesium-dependent variants. | the adeno-associated virus type 2 (aav) replication (rep) proteins rep78 and 68 (rep78/68) exhibit a number of biochemical activities required for aav replication, including specific binding to a 22-bp region of the terminal repeat, site-specific endonuclease activity, and helicase activity. individual and clusters of charged amino acids were converted to alanines in an effort to generate a collection of conditionally defective rep78/68 proteins. rep78 variants were expressed in human 293 cells ... | 1999 | 10516052 |
| repeated delivery of adeno-associated virus vectors to the rabbit airway. | efficient local expression from recombinant adeno-associated virus (raav)-cystic fibrosis (cf) transmembrane conductance regulator (cftr) vectors has been observed in the airways of rabbits and monkeys for up to 6 months following a single bronchoscopic delivery. however, it is likely that repeated administrations of raav vectors will be necessary for sustained correction of the cf defect in the airways. the current study was designed to test the feasibility of repeated airway delivery of raav v ... | 1999 | 10516053 |
| concatamerization of adeno-associated virus circular genomes occurs through intermolecular recombination. | long-term recombinant aav (raav) transgene expression in muscle has been associated with the molecular conversion of single-stranded raav genomes to high-molecular-weight head-to-tail circular concatamers. however, the mechanisms by which these large multimeric concatamers form remain to be defined. to this end, we tested whether concatamerization of raav circular intermediates occurs through intra- or intermolecular mechanisms of amplification. coinfection of the tibialis muscle of mice with ra ... | 1999 | 10516055 |
| persistent expression of canine factor ix in hemophilia b canines. | we previously demonstrated that direct intramuscular injection of recombinant adeno-associated virus (raav) carrying the human fix (hfix) cdna can safely be administered to hemophilic b canines and express human factor ix protein; however, the functional activity of the hfix protein could not be assessed due to anti-human fix antibody (inhibitor) formation. to test the therapeutic efficacy of raav in hemophilic dogs, raav type 2 (raav2) carrying canine fix (cfix) cdna was injected into the skele ... | 1999 | 10516718 |
| binding of the human papillomavirus type 16 p97 promoter by the adeno-associated virus rep78 major regulatory protein correlates with inhibition. | human papillomavirus type 16 (hpv-16) infection is positively associated with cervical cancer, whereas adeno-associated virus (aav) infection is negatively associated with this same cancer. in earlier studies these two virus types have been shown to directly interact, with aav inhibiting or enhancing papillomavirus functions depending upon the specific circumstances. one defined interaction between these two viruses is the ability of the aav rep78 major regulatory protein to inhibit gene express ... | 1999 | 10531369 |
| superior gene transfer into solid tumour cells than into human mobilised peripheral blood progenitor cells using helpervirus-free adeno-associated viral vector stocks. | autologous peripheral blood progenitor cell (pbpc) grafts can be contaminated with tumour cells that potentially give rise to relapse following myeloablative therapy and pbpc transplantation. adeno-associated virus (aav)-based vectors produced by a new adenovirus-free technique are a gene delivery system which may be applicable for tumour cell purging. to test for the host range of these vectors, solid tumours of clinical relevance and normal cd34+ pbpc were selected as target cells for an aav-v ... | 1999 | 10533460 |
| adeno-associated virus (aav) site-specific integration: formation of aav-aavs1 junctions in an in vitro system. | an in vitro system to study the mechanism of site-specific integration of adeno-associated virus (aav) was developed. this system is based on two substrates, a linear or circular aav donor and a circular acceptor containing the preintegration locus aavs1. in the presence of hela extract and the his-tag-purified rep68 protein, specific covalent junctions between aav and aavs1 were formed and detected by pcr. the majority of the junctions were located within the rep binding site of both the aav an ... | 1999 | 10536011 |
| haemophilia b gene therapy using adeno-associated virus. | | 1999 | 10542387 |
| gene transfer to neurons with herpes simplex virus/adeno-associated virus hybrid vectors. | | 1999 | 10543609 |
| progress in adeno-associated virus type 2 vector production: promises and prospects for clinical use. | vectors derived from the human parvovirus aav-2 (adeno-associated virus type 2) are among the most promising gene delivery vehicles currently being developed. these vectors are not only capable of transducing a large variety of human cell types in vitro and in vivo, but in immunocompetent animal models can establish long-term gene expression without being pathogenic to the recipient. however, a limitation of this vector system with respect to its clinical application has long been the laborious ... | 1999 | 10543610 |
| gene transfer to the nigrostriatal system by hybrid herpes simplex virus/adeno-associated virus amplicon vectors. | to improve gene transfer to cns neurons, critical elements of herpes simplex virus 1 (hsv-1) amplicons and recombinant adeno-associated virus (aav) vectors were combined to construct a hybrid amplicon vector, and then packaged via a helper virus-free system. we tested the hsv/aav hybrid amplicon vectors for transduction efficiency and stability of transgene expression (green fluorescent protein) in primary neuronal cultures from rat fetal ventral mesencephalon, in comparison with traditional hsv ... | 1999 | 10543613 |
| high-titer recombinant adeno-associated virus production from replicating amplicons and herpes vectors deleted for glycoprotein h. | production of high-titer raav is essential for in vivo clinical application. one limiting factor may be the failure of existing systems to replicate the packaging genome in such a way that expression of rep and cap proteins is coordinately amplified. disc-hsv (disabled single-cycle virus) is a genetically modified herpes simplex virus (hsv) that by deletion of glycoprotein h (gh) is infectious only if propagated in a complementing cell line. in this study, we have used disc-hsv as a helper for r ... | 1999 | 10543617 |
| infection with adeno-associated virus may protect against excitotoxicity. | gene therapy has developed as a promising approach for therapy in a broad variety of conditions. viral vectors have been developed that may replace a defective gene, prevent expression of a mutant gene, or deliver a protective gene and thereby delay cellular loss. using adeno-associated virus containing green fluorescent protein (aav-gfp) we were able to specifically transduce cells located in the inner retina and induce over-expression of gfp in adult rat retinae. the delivery and expression of ... | 1999 | 10549791 |
| overexpression of cyclin a inhibits augmentation of recombinant adeno-associated virus transduction by the adenovirus e4orf6 protein. | the 34-kda product of adenovirus e4 region open reading frame 6 (e4orf6) dramatically enhances transduction by recombinant adeno-associated virus vectors (raav). this is achieved by promoting the conversion of incoming single-stranded viral genomes into transcriptionally competent duplex molecules. the molecular mechanism for enhancing second-strand synthesis is not fully understood. in this study, we analyzed the cellular consequences of e4orf6 expression and the requirements for efficient raav ... | 1999 | 10559315 |
| dynamin is required for recombinant adeno-associated virus type 2 infection. | recombinant adeno-associated virus (raav) vectors for gene therapy of inherited disorders have demonstrated considerable potential for molecular medicine. recent identification of the viral receptor and coreceptors for aav type 2 (aav-2) has begun to explain why certain organs may demonstrate higher efficiencies of gene transfer with this vector. however, the mechanisms by which aav-2 enters cells remain unknown. in the present report, we have examined whether the endocytic pathways of raav-2 ar ... | 1999 | 10559355 |
| detection of adeno-associated virus in human semen: does viral infection play a role in the pathogenesis of male infertility? | to evaluate the occurrence of adeno-associated virus (aav) dna and/or human papillomavirus (hpv) dna in the semen of infertile men as a possible factor in the pathogenesis of male infertility. | 1999 | 10560983 |
| site-specific targeting of dna plasmids to chromosome 19 using aav cis and trans sequences. | | 2000 | 10561835 |
| adeno-associated virus based gene therapy in skeletal muscle. | | 2000 | 10561836 |
| integrin alphavbeta5 is not involved in adeno-associated virus type 2 (aav2) infection. | alphavbeta5 integrin was recently proposed as a coreceptor for adeno-associated virus type 2 (aav2) infection (summerford et al., 1999, nat. med. 5, 78-82), based mainly on the direct binding of aav2 to denatured beta5 by virus overlay assay. in studies using purified natural or recombinant human integrin alphavbeta5 we were unable to demonstrate aav2 binding, either by virus overlay or by liquid binding assay. furthermore, neither purified integrin alphavbeta5, nor rgd peptides, nor functional ... | 1999 | 10562505 |
| delayed expression of adeno-associated virus vector dna. | two previous reports indicated that recombinant adeno-associated virus (raav) vectors were dependent on helper adenovirus (ad) for efficient conversion of single-stranded (ss) raav dna to the double-stranded (ds) form. this finding is somewhat paradoxical, however, since during a latent infection wild-type (wt)-aav is rapidly converted to a ds form in the absence of ad. our hypothesis was that the effect observed in the previous studies was due to kinetic factors, i.e. to a relative delay in con ... | 1999 | 10567839 |
| a helper virus-free packaging system for recombinant adeno-associated virus vectors. | adeno-associated virus (aav) is a human parvovirus that is currently receiving widespread attention for its potential use as a gene therapy vector. construction of the recombinant aav vector (raav) involves replacing most of the viral genome with a transgene of interest and then packaging this recombinant genome into an infectious virion. most current protocols for generating raav entail the co-transfection of a vector plasmid and a packaging plasmid that expresses the viral replication and stru ... | 1999 | 10570967 |
| prevention of 6-hydroxydopamine-induced rotational behavior by bdnf somatic gene transfer. | brain-derived neurotrophic factor (bdnf) was expressed via injection of viral vector into the substantia nigra pars compacta (snc) to investigate its influence on nigrostriatal dopaminergic activity and locomotor behavior. the recombinant adeno-associated virus (raav) vector, ptr-bdnfmyc, incorporated the neuron-specific enolase (nse) promoter and the internal ribosome entry site (ires) element driving expression of both epitope-tagged bdnf and green fluorescent protein (gfp) bicistronically. th ... | 1999 | 10575102 |
| cochlear gene therapy: current perspectives. | gene therapy has the potential to revolutionize therapy for hearing disorders. advances in our understanding of the molecular basis of hearing combined with techniques for intracochlear gene transfer are leading us closer to clinical applications. potential areas of interest are reviewed and experimental success using adeno-associated virus to effect intracochlear gene transfer is discussed. | 1999 | 10577770 |
| targets for gene therapy of vein grafts. | poor long-term patency and a lack of suitable systemic pharmacologic therapy for the prevention of vein graft failure have prompted the search for effective local gene therapy. vein grafts are particularly well suited for gene transfer in the clinic because direct access to vein is available during surgical preparation for grafting. in this review, the available animal models are discussed and a new mouse model is highlighted. recent advances in gene transfer technology are reviewed, including t ... | 1999 | 10579065 |
| stable transfection of rat preporinsulin ii gene into rat hematopoietic stem cells via recombinant adeno-associated virus. | we investigated the ability of recombinant adeno-associated virus (raav), to mediate the transfer of rat preproinsulin ii (ri2) gene into rat hematopoietic stem cells in vitro and expression of ri2 following intra-venous (i.v.) injection of infected stem cells into syngeneic rats. the plp-1 recombinant plasmid containing ri2 was engineered as follows: ri2 with rsv-promoter was released from pbc 12bi (atcc), purified, and inserted into bamh1 site of raav vector plasmid pwp-19. plasmid plp-1, toge ... | 1999 | 10579458 |
| neuronal-specific and nerve growth factor-inducible expression directed by the preprotachykinin-a promoter delivered by an adeno-associated virus vector. | the ability to manipulate the expression of genes within neurons provides unique opportunities to study the role of individual gene products in nervous system function. virus vectors are a potentially rapid tool for the experimental manipulation of gene expression in the mammalian nervous system. however, a block to the use of virus vector systems in neurobiology is often the lack of cell-specific expression of the gene within the nervous system, and the immune and inflammatory responses to both ... | 1999 | 10579592 |
| genetic capsid modifications allow efficient re-targeting of adeno-associated virus type 2. | | 1999 | 10581091 |
| gene therapy of genetic diseases and cancer. | the scope of gene transfer applications in human therapy has expanded enormously over the last 15 years to include not only several types of genetic diseases but also a variety of genetic approaches to the treatment of cancer. hematopoietic stem cells have been considered excellent targets for therapeutic gene transfer because of their capacity for self-renewal and for differentiation into multiple cellular lineages. retrovirus-mediated gene transfer has been tested for treatment of diseases tha ... | 1999 | 10587981 |
| conditional site-specific integration into human chromosome 19 by using a ligand-dependent chimeric adeno-associated virus/rep protein. | it is of great interest for gene therapy to develop vectors that drive the insertion of a therapeutic gene into a chosen specific site on the cellular genome. adeno-associated virus (aav) is unique among mammalian viruses in that it integrates into a distinct region of human chromosome 19 (integration site aavs1). the inverted terminal repeats (itrs) flanking the aav genome and the aav-encoded nonstructural proteins rep78 and/or rep68 are the only viral elements necessary and sufficient for site ... | 2000 | 10590116 |
| incorporation of adeno-associated virus in a calcium phosphate coprecipitate improves gene transfer to airway epithelia in vitro and in vivo. | adeno-associated virus (aav) is inefficient at infecting differentiated airway epithelia because of a lack of receptors at the apical surface. we hypothesized that incorporation of aav in a calcium phosphate coprecipitate would circumvent this barrier. interestingly, coprecipitation of aav type 2 improved gene transfer to differentiated human airway epithelia in vitro and to the mouse lung in vivo. these results suggest that delivery of aav as a cap(i) coprecipitate may significantly enhance its ... | 2000 | 10590145 |
| chimeric virus-like particle formation of adeno-associated virus. | adeno-associated virus (aav) capsids are composed of three proteins, vp1, vp2, and vp3. these capsid proteins have a common amino acid sequence, being expressed from different initiation codons on the same open reading frame. although vp1 is necessary for viral infection, it is not essential for capsid formation. the other capsid proteins, vp2 and vp3, are sufficient for capsid formation, but their functions are poorly understood. to investigate the role(s) of the capsid proteins in capsid forma ... | 1999 | 10600510 |
| insertional mutagenesis of aav2 capsid and the production of recombinant virus. | the structural genes of adeno-associated virus serotype 2 (aav2) have been altered by linker insertional mutagenesis in order to define critical components of virion assembly and infectivity. an in-frame restriction site linker was inserted across the capsid coding domain of a recombinant plasmid. after complementation in vivo, recombinant aav2 viruses were generated and assayed for capsid production, packaging, transduction, heparin agarose binding, and morphology. three classes of capsid mutan ... | 1999 | 10600599 |
| intravenous angiotensinogen antisense in aav-based vector decreases hypertension. | angiotensinogen (agt) has been linked to hypertension. because there are no direct inhibitors of agt, we have developed antisense (as) inhibition of agt mrna delivered in an adeno-associated virus (aav)-based plasmid vector. this plasmid, driven by the cytomegalovirus promoter, contains a green fluorescent protein reporter gene and as cdna for rat agt. transfection of the plasmid into rat hepatoma cells brought a strong expression of the transgenes and a significant reduction in the level of agt ... | 1999 | 10600860 |
| adeno-associated virus-mediated gene transfer of factor ix for treatment of hemophilia b by gene therapy. | | 1999 | 10605748 |
| adeno-associated virus type 2 nonstructural protein rep78 suppresses translation in vitro. | adeno-associated virus type 2 nonstructural protein rep78 [621 amino acids (aa) long] affects the expression of various cellular and viral genes. in this study we examined the effects of rep78 on expression of the luciferase gene from the human cytomegalovirus immediate-early promoter in hela cells and on translation of rna encoding luciferase in rabbit reticulocyte lysate. when rep78 and luciferase were coexpressed, the luciferase activity decreased despite increased levels of luciferase mrna i ... | 2000 | 10612674 |
| adenoviral and adeno-associated viral transfer of genes to the peripheral nervous system. | targeted expression of foreign genes to the peripheral nervous system is interesting for many applications, including gene therapy of neuromuscular diseases, neuroanatomical studies, and elucidation of mechanisms of axonal flow. here we describe a microneurosurgical technique for injection of replication-defective viral vectors into dorsal root ganglia (drg). adenovirus- and adeno-associated virus-based vectors with transcriptional competence for drg neurons led to expression of the gene of inte ... | 2000 | 10618437 |
| gene transfer into hypothalamic organotypic cultures using an adeno-associated virus vector. | organotypic cultures of rat hypothalamic slice cultures were successfully transduced using adeno-associated viral vectors. using nuclear-targeted lac-z as the reporter gene, transduction was found to be very effective, occurring in as high as 89% of a specific cell type, the oxytocin neurons, present in the cultured explants. these transduction levels were not accompanied by any deleterious effects in the cultured cells 7 days after transduction. such an in vitro approach should be valuable for ... | 1999 | 10619549 |
| a role for single-stranded templates in cell-free adeno-associated virus dna replication. | assays have been described in which duplex adeno-associated virus (aav) dna can be replicated in hela cell extracts with exogenous aav rep protein. these assays appear to mimic the aav dna replication that occurs in the cell, including the ability of extracts from adenovirus (ad)-infected cells to replicate duplex aav dna templates more efficiently than extracts from uninfected cells can. we showed previously that the ad-infected extract was able to support a more processive replication than the ... | 2000 | 10623736 |
| impaired intracellular trafficking of adeno-associated virus type 2 vectors limits efficient transduction of murine fibroblasts. | although adeno-associated virus type 2 (aav) has gained attention as a potentially useful alternative to the more commonly used retrovirus- and adenovirus-based vectors for human gene therapy, efficient gene transfer and transgene expression by aav vectors require that the following two obstacles be overcome. first, the target cell must express the receptor and the coreceptor for aav infection, and second, the cell must allow for viral second-strand dna synthesis. we now describe a third obstacl ... | 2000 | 10623762 |
| full functional rescue of a complete muscle (ta) in dystrophic hamsters by adeno-associated virus vector-directed gene therapy. | limb girdle muscular dystrophy (lgmd) 2f is caused by mutations in the delta-sarcoglycan (sg) gene. previously, we have shown successful application of a recombinant adeno-associated virus (aav) vector for genetic and biochemical rescue in the bio14.6 hamster, a homologous animal model for lgmd 2f (j. li et al., gene ther. 6:74-82, 1999). in this report, we show efficient and long-term delta-sg expression accompanied by nearly complete recovery of physiological function deficits after a single-d ... | 2000 | 10627554 |
| repeat transduction in the mouse lung by using adeno-associated virus vectors with different serotypes. | vectors derived from adeno-associated virus type 2 (aav2) promote gene transfer and expression in the lung; however, we have found that while gene expression can persist for at least 8 months in mice, it was reduced dramatically in rabbits over a period of 2 months. the efficiency and persistence of aav2-mediated gene expression in the human lung have yet to be determined, but it seems likely that readministration will be necessary over the lifetime of an individual. unfortunately, we have found ... | 2000 | 10627564 |
| sustained production of beta-glucuronidase from localized sites after aav vector gene transfer results in widespread distribution of enzyme and reversal of lysosomal storage lesions in a large volume of brain in mucopolysaccharidosis vii mice. | the lysosomal storage disorders are a large group of inherited diseases that involve central nervous system degeneration. the disease in the brain has generally been refractory to treatment, which will require long-term correction of lesions dispersed throughout the central nervous system to be effective. a promising approach is somatic gene therapy but the methods have so far been inadequate because they have only achieved short-term or localized improvements. a potential approach to overcome t ... | 1999 | 10630187 |
| adeno-associated virus rep78 binds to e2-responsive element 1 of bovine papillomavirus type 1. | adeno-associated virus type-2 (aav-2) is a helper-dependent parvovirus that has been implicated in the inhibition of replication and oncogenic transformation of bovine papillomavirus type-1 (bpv-1) and other transforming dna viruses. previous studies have suggested that the rep78 protein of aav-2 is a key player mediating this effect. in this report we have analyzed the effect of aav-2 rep78 protein on the regulation of gene expression of a reporter gene under the control of the long control reg ... | 1999 | 10632568 |
| epitope mapping of human anti-adeno-associated virus type 2 neutralizing antibodies: implications for gene therapy and virus structure. | recombinant adeno-associated virus type 2 (aav) is a common vector used in human gene therapy protocols. we characterized the humoral immune response to aav and observed that 80% of normal human subjects have anti-aav antibodies and that 18% have neutralizing antibodies. to analyze the effect of neutralizing antibodies on aav readministration, we attempted to deliver recombinant aav expressing human factor ix (aav-hfix) intraportally into the livers of mice which had been preexposed to aav and s ... | 2000 | 10644347 |
| adeno-associated virus 2-mediated transduction and erythroid lineage-restricted expression from parvovirus b19p6 promoter in primary human hematopoietic progenitor cells. | human parvovirus b19 gene expression from the viral p6 promoter (b19p6) is restricted to primary human hematopoietic cells undergoing erythroid differentiation. we have demonstrated that expression from this promoter does not occur in established human erythroid cell lines in the context of a recombinant parvovirus genome (ponnazhagan et al. j virol 69:8096-8101, 1995). however, abundant expression from this promoter can be readily detected in primary human bone marrow cells (wang et al. proc na ... | 1999 | 10645765 |
| viral vectors for gene delivery and gene therapy within the endocrine system. | the transfer of genetic material into endocrine cells and tissues, both in vitro and in vivo, has been identified as critical for the study of endocrine mechanisms and the future treatment of endocrine disorders. classical methods of gene transfer, such as transfection, are inefficient and limited mainly to delivery into actively proliferating cells in vitro. the development of viral vector gene delivery systems is beginning to circumvent these initial setbacks. several kinds of viruses, includi ... | 2000 | 10657846 |
| a chimeric protein containing the n terminus of the adeno-associated virus rep protein recognizes its target site in an in vivo assay. | the rep78 and rep68 proteins of adeno-associated virus (aav) type 2 are involved in dna replication, regulation of gene expression, and targeting site-specific integration. they bind to a specific rep recognition sequence (rrs) found in both the viral inverted terminal repeats and the aavs1 integration locus on human chromosome 19. previous in vitro studies implied that an n-terminal segment of rep is involved in dna recognition, while additional domains might stabilize binding and mediate multi ... | 2000 | 10666268 |
| humoral immunity to adeno-associated virus type 2 vectors following administration to murine and nonhuman primate muscle. | adeno-associated virus (aav) is being developed as a vector capable of conferring long-term gene expression, which is useful in the treatment of chronic diseases. in most therapeutic applications, it is necessary to readminister the vector. this study characterizes the humoral immune response to aav capsid proteins following intramuscular injection and its impact on vector readministration. studies of mice and rhesus monkeys demonstrated the formation of neutralizing antibodies to aav capsid pro ... | 2000 | 10666273 |
| adeno-associated virus major rep78 protein disrupts binding of tata-binding protein to the p97 promoter of human papillomavirus type 16. | adeno-associated virus type 2 (aav) is known to inhibit the promoter activities of several oncogenes and viral genes, including the human papillomavirus type 16 (hpv-16) e6 and e7 transforming genes. however, the target elements of aav on the long control region (lcr) upstream of e6 and e7 oncogenes are elusive. a chloramphenicol acetyltransferase assay was performed to study the effect of aav on the transcription activity of the hpv-16 lcr in siha (hpv-positive) and c-33a (hpv-negative) cells. ... | 2000 | 10666281 |
| transduction and utility of the granulocyte-macrophage colony-stimulating factor gene into monocytes and dendritic cells by adeno-associated virus. | the genetic manipulation of antigen-presenting dendritic cells (dc) offers promise for stimulating the immune response, in particular for anticancer and antiviral protocols. as adeno-associated virus (aav) has shown promise as a gene delivery vector for transducing a variety of hematopoietic cell types, we have investigated aav's ability to genetically alter dc. in this analysis, we modified the standard granulocyte-macrophage colony-stimulating factor (gm-csf) and interleukin-4 (il-4) treatment ... | 2000 | 10670649 |
| developments in gene therapy for muscular dystrophy. | gene therapy for muscular dystrophy (md) presents significant challenges, including the large amount of muscle tissue in the body, the large size of many genes defective in different muscular dystrophies, and the possibility of a host immune response against the therapeutic gene. overcoming these challenges requires the development and delivery of suitable gene transfer vectors. encouraging progress has been made in modifying adenovirus (ad) vectors to reduce immune response and increase capacit ... | 2000 | 10679969 |
| aav vectors: is clinical success on the horizon? | potential applications and impact of the adeno-associated virus (aav) as a gene transfer vector have expanded rapidly in the last decade. recent advances in the production of high-titer purified raav vector stocks have made the transition to human clinical trials a reality in the last moments of the millenium. production improvements will be complemented in the coming years with understanding of and innovations in the targeting and packaging of raav, the design of transgene cassettes, and the ho ... | 2000 | 10680012 |
| gene transfer into the cns using recombinant adeno-associated virus: analysis of vector dna forms resulting in sustained expression. | recombinant adeno-associated virus (raav) vectors have shown significant promise as vehicles for in vivo gene transfer, particularly for transduction of organs composed primarily of non-dividing cells (i.e., muscle, cns, and liver). however, the mechanistic basis for this desirable property remains unclear. to investigate the fate of raav genomes in mouse brain, we stereotactically injected an raav vector carrying the e. coli lacz gene into the caudate of balb/c mice and demonstrate efficient tr ... | 1999 | 10682906 |
| loss of atm function enhances recombinant adeno-associated virus transduction and integration through pathways similar to uv irradiation. | ataxia telangiectasia is caused by a genetic defect in the atm gene that results in altered cellular sensitivity to dna-damaging agents such as gamma-irradiation. atm deficiency is associated with an increased incidence of neurological disorders, immune deficiency, and cancer. in this report we demonstrate that recombinant adeno-associated virus (raav) gene transfer in atm-deficient fibroblasts is significantly enhanced over normal fibroblast cell lines. this enhancement of raav transduction in ... | 2000 | 10683328 |
| infectious entry pathway of adeno-associated virus and adeno-associated virus vectors. | we have investigated the infectious entry pathway of adeno-associated virus (aav) and recombinant aav vectors by assessing aav-mediated gene transfer and by covalently conjugating fluorophores to aav and monitoring entry by fluorescence microscopy. we examined aav entry in hela cells and in hela cell lines which inducibly expressed a dominant interfering mutant of dynamin. the data demonstrate that aav internalizes rapidly by standard receptor-mediated endocytosis from clathrin-coated pits (half ... | 2000 | 10684294 |
| recombinant adeno-associated virus expressing human papillomavirus type 16 e7 peptide dna fused with heat shock protein dna as a potential vaccine for cervical cancer. | in this study, we explore a potential vaccine for human papillomavirus (hpv)-induced tumors, using heat shock protein as an adjuvant, a peptide vaccine for safety, and adeno-associated virus (aav) as a gene delivery vector. the tumor vaccine was devised by constructing a chimeric gene which contained hpv type 16 e7 cytotoxic t-lymphocyte (ctl) epitope dna (m. c. feltkamp, h. l. smits, m. p. vierboom, r. p. minnaar, b. m. de jongh, j. w. drijfhout, j. ter schegget, c. j. melief, and w. m. kast, e ... | 2000 | 10684306 |
| mutational analysis of adeno-associated virus type 2 rep68 protein endonuclease activity on partially single-stranded substrates. | the endonuclease activity of the rep68 and rep78 proteins (rep68/78) of adeno-associated virus type 2 (aav) cuts at the terminal resolution site (trs) within the hairpin structure formed by the aav inverted terminal repeats. recent studies suggest that a dna unwinding function of rep68/78 may be required for endonuclease activity. we demonstrate that several mutant proteins which are endonuclease negative on a fully duplex hairpin substrate are endonuclease positive on a partially single-strande ... | 2000 | 10684315 |
| an oral vaccine against nmdar1 with efficacy in experimental stroke and epilepsy. | the brain is generally considered immunoprivileged, although increasing examples of immunological responses to brain antigens, neuronal expression of major histocompatibility class i genes, and neurological autoimmunity have been recognized. an adeno-associated virus (aav) vaccine generated autoantibodies that targeted a specific brain protein, the nr1 subunit of the n-methyl-d-aspartate (nmda) receptor. after peroral administration of the aav vaccine, transgene expression persisted for at least ... | 2000 | 10688787 |
| sustained expression of human factor viii in mice using a parvovirus-based vector. | persistent therapeutic levels of human factor viii (hfviii) would signify a major advance in the treatment of hemophilia a. here we report sustained expression of hfviii in immunocompetent mice using recombinant adeno-associated virus (raav) vectors. aav can stably transduce liver cells, the target tissue for efficient hfviii production. because of raav packaging constraints, we tested 2 constructs using small regulatory elements designed for liver-specific transgene expression linked to b-domai ... | 2000 | 10688813 |
| recombinant adeno-associated virus type 2, 4, and 5 vectors: transduction of variant cell types and regions in the mammalian central nervous system. | recombinant adeno-associated virus vectors based on serotype 2 (raav2) can direct transgene expression in the central nervous system (cns), but it is not known how other raav serotypes perform as cns gene transfer vectors. serotypes 4 and 5 are distinct from raav2 and from each other in their capsid regions, suggesting that they may direct binding and entry into different cell types. in this study, we examined the tropisms and transduction efficiencies of beta-galactosidase-encoding vectors made ... | 2000 | 10688913 |
| therapy and prevention of arthritis by recombinant adeno-associated virus vector with delivery of interleukin-1 receptor antagonist. | to evaluate the recombinant adeno-associated virus vector encoding interleukin-1 receptor antagonist (raav-il-1ra) complementary dna for its potential in the treatment and prevention of lipopolysaccharide (lps)-induced arthritis. | 2000 | 10693868 |
| additional transduction events after subretinal readministration of recombinant adeno-associated virus. | subretinal delivery of recombinant adeno-associated virus (raav) results in a systemic humoral response in the adult immunocompetent mouse. we characterized this response and determined whether it is possible to readminister raav to the subretinal space despite the presence of antibodies to the vector. a systemic humoral response to raav capsid proteins was induced by either unilateral subretinal injection or by intradermal administration of 1 x 10(9) infectious units of raav carrying the cdna e ... | 2000 | 10697119 |
| site-specific integration of adeno-associated virus-based plasmid vectors in lipofected hela cells. | adeno-associated virus (aav) integrates specifically into a site (aavs1) on human chromosome 19q13.3-qter. similarly, there is accumulating evidence that this site-specific integration occurs by transfection of aav-based plasmid vectors. in order to further define the process of plasmid integration events, we constructed some aav plasmids, introduced them into hela cells by lipofection, and isolated chromosomal integrants. one of such plasmids, pth-5, contained the rep and neomycin-resistant (ne ... | 2000 | 10704347 |
| an adeno-associated virus (aav) initiator protein, rep78, catalyzes the cleavage and ligation of single-stranded aav ori dna. | the rep78 protein of adeno-associated virus (aav) contains amino acid sequence motifs common to rolling-circle replication (rcr) initiator proteins. in this report, we describe rcr initiator-like activities of rep78. we demonstrate that a maltose-binding protein (mbp)-rep78 fusion protein can catalyze the cleavage and ligation of single-stranded dna substrates derived from the aav origin of replication. rep-mediated single-stranded dna cleavage was strictly dependent on the presence of certain d ... | 2000 | 10708427 |
| targeting the urokinase plasminogen activator receptor enhances gene transfer to human airway epithelia. | developing gene therapy for cystic fibrosis has been hindered by limited binding and endocytosis of vectors by human airway epithelia. here we show that the apical membrane of airway epithelia express the urokinase plasminogen activator receptor (upar). urokinase plasminogen activator (upa), or a 7-residue peptide derived from this protein (u7-peptide), bound the receptor and stimulated apical endocytosis. both ligands enhanced gene transfer by nonspecifically bound adenovirus and adeno-associat ... | 2000 | 10712430 |
| a method for the preparation of highly purified adeno-associated virus using affinity column chromatography, protease digestion and solvent extraction. | recombinant adeno-associated virus (aav) is becoming the vector of choice for many gene therapy protocols. there has been much recent progress made toward increasing aav titres but a continuing problem in using aav has been that it is relatively difficult to concentrate and purify. traditional methods, such as caesium chloride (cscl) gradients, have drawbacks, notably extended purification times and the ability to process only limited volumes. where the target cells of interest require a high mu ... | 2000 | 10716335 |
| effect of tt virus infection on hepatocellular carcinoma development: results of a euro-asian survey. | a small percentage of persons with hepatocellular carcinoma (hcc) lack identifiable causes of liver pathology. the single-stranded dna virus, tt virus (ttv), has been found in persons with acute and chronic liver injury. nested polymerase chain reaction was used to search for both ttv and parvoviruses in 293 hcc samples from asia and europe. ttv was found in >30% of chinese and italian samples but in only 13% of french samples. no clinicopathologic differences were found between ttv-positive and ... | 2000 | 10720542 |
| oligodendrocyte-specific gene expression in mouse brain: use of a myelin-forming cell type-specific promoter in an adeno-associated virus. | to explore the feasibility of cell type-specific gene expression in oligodendrocytes as a possible therapeutic approach for demyelinating diseases, the cell specificity, tissue specificity, and duration of gene expression were investigated using recombinant adeno-associated viral vectors (raav) carrying a green fluorescence protein (gfp) gene. recombinant aav vectors carrying either the myelin basic protein (mbp) promoter (raav-mbp-gfp) or the cytomegalovirus (cmv) immediate early promoter (raav ... | 1999 | 10723060 |
| recombinant adeno-associated virus-mediated correction of lysosomal storage within the central nervous system of the adult mucopolysaccharidosis type vii mouse. | the central nervous system (cns) is a predominant site of involvement in several lysosomal storage diseases (lsds); and for many patients, these diseases are diagnosed only after the onset of symptoms related to the progressive accumulation of macromolecules within lysosomes. the mucopolysaccharidosis type vii (mps vii) mice are deficient for the lysosomal enzyme beta-glucuronidase and, by early adulthood, develop a significant degree of glycosaminoglycan storage within neuronal, glial, and lept ... | 2000 | 10724030 |
| the use of adeno-associated virus to circumvent the maturation-dependent viral transduction of muscle fibers. | muscle-based gene therapy using adenovirus, retrovirus, and herpes simplex virus has been hindered by viral cytotoxicity, host immune response, and the maturation-dependent viral transduction of muscle fibers. the development of new mutant vectors has greatly reduced the toxicity and the immune rejection problems, but the inability of viral vectors to penetrate and transduce mature myofibers remains an important issue. research has been focused on the characterization of barriers to viral transd ... | 2000 | 10724031 |
| adeno-associated virus vectors show variable dependence on divalent cations for thermostability: implications for purification and handling. | recombinant adeno-associated virus (raav) shows significant promise as a vector for gene transfer in pre-clinical models of human disease, and is currently being evaluated in human clinical trials. as a consequence, increasing attention is being turned to the important tasks of optimizing raav titer, purity, and stability. we have observed dramatic variation in divalent cation dependence for thermostability of different raav vectors. to further investigate this observation, the thermostability o ... | 2000 | 10724041 |
| the interaction of heparin sulfate and adeno-associated virus 2. | recently heparan sulfate was proposed as the host cell receptor for the dependovirus, adeno-associated virus type 2 (aav2). we show that although heparan sulfate on the cell surface may contribute to the binding of aav2 to permissive cells, the amount of heparan sulfate on the cell surface as determined by flow cytometry using four different monoclonal antibodies does not correlate with aav2 binding to cells or recombinant aav2 transduction efficiency. experiments with either mutant cho cells or ... | 2000 | 10725206 |
| productive replication of adeno-associated virus can occur in human papillomavirus type 16 (hpv-16) episome-containing keratinocytes and is augmented by the hpv-16 e2 protein. | we used a sensitive assay to test whether an adeno-associated virus (aav) productive replication cycle can occur in immortalized human keratinocytes carrying episomal human papillomavirus type 16 (hpv-16) dna. following transfection with cloned aav dna, infectious aav was produced, and the infectivity was blocked by anti-aav antiserum. the hpv-16 e2 protein substantially increased the yield of aav. other hpv early proteins did not, in our experiments, show this ability. e2 has been shown to be a ... | 2000 | 10729123 |
| kinetics of recombinant adeno-associated virus-mediated gene transfer. | recombinant adeno-associated virus (raav) vectors have been shown to be useful for efficient gene delivery to a variety of dividing and nondividing cells. mechanisms responsible for the long-term, persistent expression of the raav transgene are not well understood. in this study we investigated the kinetics of raav-mediated human factor ix (hfix) gene transfer into human primary myoblasts and myotubes. transduction of both myoblasts and myotubes occured with a similar and high efficiency. after ... | 2000 | 10729130 |
| nonrandom transduction of recombinant adeno-associated virus vectors in mouse hepatocytes in vivo: cell cycling does not influence hepatocyte transduction. | recombinant adeno-associated virus vectors (raav) show promise in preclinical trials for the treatment of genetic diseases including hemophilia. liver-directed gene transfer results in a slow rise in transgene expression, reaching steady-state levels over a period of 5 weeks concomitant with the conversion of the single-stranded raav molecules into high-molecular-weight concatemers in about 5% of hepatocytes. immunohistochemistry and rna in situ hybridization show that the transgene product is m ... | 2000 | 10729154 |
| adeno-associated virus type 5 (aav5) but not aav2 binds to the apical surfaces of airway epithelia and facilitates gene transfer. | in the genetic disease cystic fibrosis, recombinant adeno-associated virus type 2 (aav2) is being investigated as a vector to transfer cftr cdna to airway epithelia. however, earlier work has shown that the apical surface of human airway epithelia is resistant to infection by aav2, presumably as a result of a lack of heparan sulfate proteoglycans on the apical surface. this inefficiency can be overcome by increasing the amount of vector or by increasing the incubation time. however, these interv ... | 2000 | 10729159 |