Publications
| Title | Abstract | Year Filter | PMID(sorted descending) Filter |
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| chromosomal latency and expression at map unit 96 of a wild-type plus adeno-associated virus (aav)/neo vector and identification of p81, a new aav transcriptional promoter. | human adeno-associated virus (aav) is ubiquitous and known to establish latency by chromosomal integration. we have constructed a wild-type plus aav vector, ins96-0.9neo, containing the neomycin resistance gene open reading frame (neo orf) of 960 bases in length at map unit 96 of the virus. ins96-0.9neo was constructed in an unconventional manner in that the neo orf lacked a dedicated heterologous promoter. in this study, this wild-type plus aav vector was to used to test aav's packaging capacit ... | 1999 | 10774553 |
| adeno-associated virus site-specifically integrates into a muscle-specific dna region. | the nonpathogenic human virus adeno-associated virus type 2 (aav) has evolved the potentially unique strategy to establish latency by site-specifically integrating its genome into human chromosome 19 (19q13.3-qter) at a locus designated aavs1. this nonhomologous, site-specific recombination of viral dna with the human genome provides a basis for developing targeted gene therapy vectors. to assess whether the region surrounding aavs1 might have contributed to the selection of the specific integra ... | 2000 | 10758163 |
| increased motoneuron survival and improved neuromuscular function in transgenic als mice after intraspinal injection of an adeno-associated virus encoding bcl-2. | mutations in the gene encoding cu/zn superoxide dismutase (sod1) underlie some familial cases of amyotrophic lateral sclerosis (als), a neurodegenerative disorder characterized by loss of cortical, brainstem and spinal motoneurons. transgenic mice over- expressing a mutated form of human sod1 containing a gly-->ala substitution at position 93 (sod1(g93a)) develop a severe, progressive motoneuron disease. we investigated the potential of recombinant adeno-associated virus (raav) to transfer neuro ... | 2000 | 10749988 |
| 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 |
| 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 |
| 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 |
| 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 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| repeat transduction in the mouse lung by using adeno-associated virus vectors with different serotypes. | vectors derived from adeno-associated virus type 2 (aav2) promote gene transfer and expression in the lung; however, we have found that while gene expression can persist for at least 8 months in mice, it was reduced dramatically in rabbits over a period of 2 months. the efficiency and persistence of aav2-mediated gene expression in the human lung have yet to be determined, but it seems likely that readministration will be necessary over the lifetime of an individual. unfortunately, we have found ... | 2000 | 10627564 |
| adeno-associated virus type 2 nonstructural protein rep78 suppresses translation in vitro. | adeno-associated virus type 2 nonstructural protein rep78 [621 amino acids (aa) long] affects the expression of various cellular and viral genes. in this study we examined the effects of rep78 on expression of the luciferase gene from the human cytomegalovirus immediate-early promoter in hela cells and on translation of rna encoding luciferase in rabbit reticulocyte lysate. when rep78 and luciferase were coexpressed, the luciferase activity decreased despite increased levels of luciferase mrna i ... | 2000 | 10612674 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| gene transfer into the mammalian inner ear using hsv-1 and vaccinia virus vectors. | the introduction of foreign genes into cells has become an effective means of achieving intracellular expression of foreign proteins, both for therapeutic purposes and for experimental manipulation. gene delivery to the nervous system has been extensively studied, primarily using viral vectors. however, to date less work has focused on gene delivery to the inner ear, and existing studies have primarily used adenovirus and adeno-associated virus. using two recombinant viral vectors, herpes simple ... | 1999 | 10452370 |
| high-fidelity correction of mutations at multiple chromosomal positions by adeno-associated virus vectors. | the gene targeting techniques used to modify chromosomes in mouse embryonic stem cells have had limited success with many other cell types, especially normal primary cells with restricted growth capacity outside the organism. this is due in large part to the technical problems and/or inefficiency of conventional dna transfer methods, as well as the low rates of homologous recombination obtained in unselected cell populations. we recently described an alternative approach in which adeno-associate ... | 1999 | 10438827 |
| efficient gene transfer into human keratinocytes with recombinant adeno-associated virus vectors. | gene transfer into the skin is a promising approach to treat inherited or acquired dermatological diseases and systemic monogenic deficiencies. for this purpose, the efficient and sustained gene delivery into keratinocytes is of critical importance. recombinant adeno-associated virus (raav) vectors hold the potential to achieve a long-term gene transfer into various human organs. in order to evaluate this potential for skin gene therapy, human keratinocytes were transduced in vitro with raav vec ... | 1999 | 10435093 |
| adeno-associated viral vectors for gene transfer and gene therapy. | adeno-associated virus (aav) is a defective, non-pathogenic human parvovirus that depends for growth on coinfection with a helper adenovirus or herpes virus. recombinant adeno-associated viruses (raavs) have attracted considerable interest as vectors for gene therapy. in contrast to other gene delivery systems, raavs lack all viral genes and show long-term gene expression in vivo without immune response or toxicity. over the past few years, many applications of raavs as therapeutic agents have d ... | 1999 | 10430026 |
| [effect of mechanical damage on ex vivo dna virus vector-mediated gene transduction in epithelial cells of murine trachea]. | the mechanism of adenovirus (ad) and adeno-associated virus (aav) vector-mediated gene transduction in murine tracheae has not been fully understood. excised tracheae from mice were exposed to either ad vector (ad-cmv-lacz) or aav vector (aav-cmv-lacz) for 1 hour. lacz gene expression in tracheal epithelial cells was detected by x-gal staining. only patch distributions of lacz expressing cells were observed. the percentage of lacz expressing cells to total cells was less than 1% with either vect ... | 1999 | 10390962 |
| position-independent human beta-globin gene expression mediated by a recombinant adeno-associated virus vector carrying the chicken beta-globin insulator. | the position-independent expression of transgenes in target cells is an essential subject for determining effective gene therapies. the chicken beta-globin insulator blocks the effects of regulatory sequences on transcriptional units at differential domains. we prepared a recombinant adeno-associated virus (raav) containing various combinations of the dnase i-hypersensitive site 2 (hs2), 3 (hs3), and 4 (hs4) core elements from the human beta-globin locus control region (lcr), the human beta-glob ... | 1999 | 10319578 |
| stable restoration of the sarcoglycan complex in dystrophic muscle perfused with histamine and a recombinant adeno-associated viral vector. | limb-girdle muscular dystrophies 2c-f represent a family of autosomal recessive diseases caused by defects in sarcoglycan genes. the cardiomyopathic hamster is a naturally occurring model for limb-girdle muscular dystrophy caused by a primary deficiency in delta-sarcoglycan. we show here that acute sarcolemmal disruption occurs in this animal model during forceful muscle contraction. a recombinant adeno-associated virus vector encoding human delta-sarcoglycan conferred efficient and stable genet ... | 1999 | 10202936 |
| targeted adeno-associated virus vector transduction of nonpermissive cells mediated by a bispecific f(ab'gamma)2 antibody. | we have developed a system for the targeted delivery of adeno-associated virus (aav) vectors. targeting is achieved via a bispecific f(ab')2 antibody that mediates a novel interaction between the aav vector and a specific cell surface receptor expressed on human megakaryocytes. targeted aav vectors were able to transduce megakaryocyte cell lines, dami and mo7e, which were nonpermissive for normal aav infection, 70-fold above background and at levels equivalent to permissive k562 cells. transduct ... | 1999 | 10052356 |
| modulation of the cytotoxicity of 3'-azido-3'-deoxythymidine and methotrexate after transduction of folate receptor cdna into human cervical carcinoma: identification of a correlation between folate receptor expression and thymidine kinase activity. | cervical carcinoma is an aids-defining illness. the expression of folate receptors (frs) in cervical carcinoma (hela-iu1) cells was modulated by stable transduction of fr cdna encapsidated in recombinant adeno-associated virus-2 in the sense and antisense orientation (sense and antisense cells, respectively). although sense cells proliferated slower than antisense or untransduced cells in vivo and in vitro in 2% (but not 10%) fcs, [methyl-3h]thymidine incorporation into dna was significantly inc ... | 1999 | 10029088 |
| development of animal models for adeno-associated virus site-specific integration. | the adeno-associated virus (aav) is unique in its ability to target viral dna integration to a defined region of human chromosome 19 (aavs1). since aavs1 sequences are not conserved in a rodent's genome, no animal model is currently available to study aav-mediated site-specific integration. we describe here the generation of transgenic rats and mice that carry the aavs1 3.5-kb dna fragment. to test the response of the transgenic animals to rep-mediated targeting, primary cultures of mouse fibrob ... | 1999 | 9971837 |
| structure of adeno-associated virus vector dna following transduction of the skeletal muscle. | the skeletal muscle provides a very permissive physiological environment for adeno-associated virus (aav) type 2-mediated gene transfer. we have studied the early steps leading to the establishment of permanent transgene expression, after injection of recombinant aav (raav) particles in the quadriceps muscle of mice. the animals received an raav encoding a secreted protein, murine erythropoietin (mepo), under the control of the human cytomegalovirus major immediate-early promoter and were sacrif ... | 1999 | 9971774 |
| reconstitution of nadph oxidase activity in human x-linked chronic granulomatous disease myeloid cells after stable gene transfer using a recombinant adeno-associated virus 2 vector. | x-linked chronic granulomatous disease (x-cgd) is an inherited disorder of host defense that results from mutations in the gene encoding gp91phox, the large subunit of the phagocyte nadph oxidase flavocytochrome b. in this study, we constructed a recombinant adeno-associated virus-2 (aav) vector in which the constitutively active promoter from the human elongation factor- 1alpha (ef-1alpha) gene drives expression of the murine gp91phox cdna, and tested its ability to integrate and express in a h ... | 1998 | 9880243 |
| viral mediated expression of insulin-like growth factor i blocks the aging-related loss of skeletal muscle function. | during the aging process, mammals lose up to a third of their skeletal muscle mass and strength. although the mechanisms underlying this loss are not entirely understood, we attempted to moderate the loss by increasing the regenerative capacity of muscle. this involved the injection of a recombinant adeno-associated virus directing overexpression of insulin-like growth factor i (igf-i) in differentiated muscle fibers. we demonstrate that the igf-i expression promotes an average increase of 15% i ... | 1998 | 9861016 |
| adeno-associated virus expression systems for gene transfer. | in contrast to other gene delivery systems, adeno-associated virus vectors show long term gene expression without immune response or toxicity. new production methods have increased vector titers and eliminated adenovirus contamination, thereby facilitating effective in vivo use. these advancements will expedite additional animal model studies providing validation for use of this vector in human clinical trials. | 1998 | 9821274 |
| adeno-associated virus as a vector for liver-directed gene therapy. | factors relevant to the successful application of adeno-associated virus (aav) vectors for liver-directed gene therapy were evaluated. vectors with different promoters driving expression of human alpha-1-antitrypsin (alpha-1at) were injected into the portal circulation of immunodeficient mice. alpha-1at expression was stable but dependent on the promoter. southern analysis of liver dna revealed approximately 0.1 to 2.0 provirus copies/diploid genome in presumed head-to-tail concatamers. in situ ... | 1998 | 9811765 |
| adeno-associated virus type 2-mediated gene transfer: role of epidermal growth factor receptor protein tyrosine kinase in transgene expression. | adeno-associated virus type 2 (aav), a single-stranded, dna-containing, nonpathogenic human parvovirus, has gained attention as a potentially useful vector for human gene therapy. however, the transduction efficiency of aav vectors varies greatly in different cells and tissues in vitro and in vivo. we have recently documented that a cellular tyrosine phosphoprotein, designated the single-stranded d-sequence-binding protein (ssd-bp), plays an important role in aav-mediated transgene expression (k ... | 1998 | 9811719 |
| successful readministration of adeno-associated virus vectors to the mouse lung requires transient immunosuppression during the initial exposure. | the airway is an important target for gene transfer to treat cystic fibrosis and other diseases that affect the lung. we previously found that marker gene expression did not persist in the bronchial epithelium following adeno-associated virus (aav) vector administration to the rabbit lung. in an attempt to promote continued expression, we tested repeat vector administration, but no additional transduction was observed, and the block to transduction correlated with the appearance of neutralizing ... | 1998 | 9811715 |
| solution structure of a na cation stabilized dna quadruplex containing g.g.g.g and g.c.g.c tetrads formed by g-g-g-c repeats observed in adeno-associated viral dna. | we have applied nmr and molecular dynamics computations including intensity based refinement to define the structure of the d(g-g-g-c-t4-g-g-g-c) dodecanucleotide in 100 mm nacl solution. the g-g-g-c sequence is of interest since it has been found as tandem repeats in the dna sequence of human chromosome 19. the same g-g-g-c sequence is also seen as islands in adeno-associated virus, a human parvovirus, which is unique amongst eukaryotic dna viruses in its ability to integrate site-specifically ... | 1998 | 9737926 |
| inhibition/stimulation of bovine papillomavirus by adeno-associated virus is time as well as multiplicity dependent. | infection by adeno-associated virus (aav) is associated with lower cervical cancer rates. we have been investigating the hypothesis that aav interacts with and inhibits the role of human papillomaviruses (hpv) in cervical cancer. we have been studying the response of bovine papillomavirus type 1 (bpv) oncogenic transformation and dna replication to aav as a prototype system. the aav rep 78 gene product is responsible for this inhibition. here, it is demonstrated that in two assay systems, focus ... | 1998 | 9705917 |
| lipofection of purified adeno-associated virus rep68 protein: toward a chromosome-targeting nonviral particle. | adeno-associated virus (aav) integrates very efficiently into a specific site (aavs1) of human chromosome 19. two elements of the aav genome are sufficient: the inverted terminal repeats (itrs) and the rep78 or rep68 protein. the incorporation of the aav integration machinery in nonviral delivery systems is of great interest for gene therapy. we demonstrate that purified recombinant rep68 protein is functionally active when directly delivered into human cells by using the polycationic liposome l ... | 1998 | 9696870 |
| adeno-associated virus gene transfer into renal cells: potential for in vivo gene delivery. | the human parvovirus adeno-associated virus (aav), type 2, has a number of features that make it an attractive choice as a vector for gene delivery to the kidney. aav vectors permit long-term gene expression in vivo by integration into the host genome, have potential for site-specific integration on chromosome 19, do not express viral genes or generate a cellular immune response, and demonstrate enhancement of gene expression by chemotherapeutic agents that are approved for use in vivo. these pr ... | 1998 | 9639033 |
| adeno-associated virus vector mediated transduction of primary normal human breast epithelial cells. | cultured human breast epithelial cells from reduction mammoplasty specimens were transduced using an adeno-associated virus vector encoding the marker gene e. coli -galactosidase. subconfluent, growing, breast epithelial cells were more easily transduced than confluent, quiescent, cells. transduction of non-dividing confluent cells could be greatly increased by ultraviolet light-induced dna damage or by prior exposure to the dna synthesis inhibitor hydroxyurea. the effects of ultraviolet light a ... | 1998 | 9625820 |
| adeno-associated viral vector-mediated gene transfer of human blood coagulation factor ix into mouse liver. | recombinant adeno-associated virus vectors (aav) were prepared in high titer (10(12) to 10(13) particles/ml) for the expression of human factor ix after in vivo transduction of murine hepatocytes. injection of aav-cmv-f.ix (expression from the human cytomegalovirus ie enhancer/promoter) into the portal vein of adult mice resulted in no detectable human factor ix in plasma, but in mice injected intravenously as newborns with the same vector, expression was initially 55 to 110 ng/ml. the expressio ... | 1998 | 9616156 |
| direct gene transfer into human epileptogenic hippocampal tissue with an adeno-associated virus vector: implications for a gene therapy approach to epilepsy. | virus vectors capable of transferring genetic information into human cells provide hope for improved therapy in several neurological diseases, including epilepsy. we evaluated the ability of an adeno-associated virus (aav) vector to transfer and cause expression of a lacz marker gene in brain slices obtained from patients undergoing temporal lobectomy for control of medically intractable seizures. | 1997 | 9579902 |
| human gene targeting by viral vectors. | stable transduction of mammalian cells typically involves random integration of viral vectors by non-homologous recombination. here we report that vectors based on adeno-associated virus (aav) can efficiently modify homologous human chromosomal target sequences. both integrated neomycin phosphotransferase genes and the hypoxanthine phosphoribosyltransferase gene were targeted by aav vectors. site-specific genetic modifications could be introduced into approximately 1% of cells, with the highest ... | 1998 | 9537413 |
| adeno-associated virus type 2-mediated gene transfer: correlation of tyrosine phosphorylation of the cellular single-stranded d sequence-binding protein with transgene expression in human cells in vitro and murine tissues in vivo. | although the adeno-associated virus type 2 (aav)-based vector system has gained attention as a potentially useful alternative to the more commonly used retroviral and adenoviral vectors for human gene therapy, the single-stranded nature of the viral genome, and consequently the rate-limiting second-strand viral dna synthesis, significantly affect its transduction efficiency. we have identified a cellular tyrosine phosphoprotein, designated the single-stranded d sequence-binding protein (ssd-bp), ... | 1998 | 9445062 |
| purification and characterization of an active form of the p78rep protein of adeno-associated virus type 2 expressed in escherichia coli. | the 78-kda product (p78rep) of the rep gene of aav-2 was expressed with an amino-terminal histidine-tag in escherichia coli and was purified under denaturing conditions. after renaturation of the p78rep protein by serial steps of dialysis, the biochemical activities of the p78rep protein were demonstrated, which include the atp-dependent endonuclease and helicase activity as well as sequence-specific binding to the aav-2 terminal repeat. these activities were retained when the protein was purifi ... | 1997 | 9425627 |
| infectious clones and vectors derived from adeno-associated virus (aav) serotypes other than aav type 2. | adeno-associated viruses (aavs) are single-stranded dependent parvoviruses being developed as transducing vectors. although at least five serotypes exist (aav types 1 to 5 [aav1 to -5]), only aav2, aav3, and aav4 have been sequenced, and the vectors in use were almost all derived from aav2. here we report the cloning and sequencing of a second aav3 genome and a new aav serotype designated aav6 that is related to aav1. aav2, aav3, and aav6 were 82% identical at the nucleotide sequence level, and ... | 1998 | 9420229 |
| midbrain injection of recombinant adeno-associated virus encoding rat glial cell line-derived neurotrophic factor protects nigral neurons in a progressive 6-hydroxydopamine-induced degeneration model of parkinson's disease in rats. | a recombinant adeno-associated virus (raav) vector capable of infecting cells and expressing rat glial cell line-derived neurotrophic factor (rgdnf), a putative central nervous system dopaminergic survival factor, under the control of a potent cytomegalovirus (cmv) immediate/early promoter (aav-md-rgdnf) was constructed. two experiments were performed to evaluate the time course of expression of raav-mediated gdnf protein expression and to test the vector in an animal model of parkinson's diseas ... | 1997 | 9391156 |
| robust, but transient expression of adeno-associated virus-transduced genes during human t lymphopoiesis. | recombinant adeno-associated viruses (raav) have been proposed to be gene transfer vehicles for hematopoietic stem cells with advantages over other virus-based systems due to their high titers and relative lack of dependence on cell cycle for target cell integration. we evaluated raav vector containing a lacz reporter gene under the control of a cytomegalovirus (cmv) promoter in the context of primary human cd34+cd2- progenitor cells induced to undergo t-cell differentiation using an in vitro t- ... | 1997 | 9389702 |
| is gene therapy in cystic fibrosis a realistic expectation? | to date there are 11 human protocols either ongoing or approved for gene therapy for cystic fibrosis (cf) in the united states. there are also two protocols in the united kingdom and one in france. of these, results have been published in four. the protocols vary in the cells targeted, the vectors used, and the frequency of administration, but despite these differences all have contributed toward answering the key questions that will determine the future of gene therapy for cf: the questions of ... | 1996 | 9363187 |
| multiple cellular proteins are recognized by the adeno-associated virus rep78 major regulatory protein and the amino-half of rep78 is required for many of these interactions. | adeno-associated virus (aav) encoded rep78 is a multi-functional protein which is required for aav dna replication, is able to regulate both aav and heterologous gene expression at the transcriptional level, and appears necessary for site specific integration of aav dna into human chromosome 19. by comparison with the analogous replication protein of the polyomaviruses, large t antigen, it seemed likely that rep78 would interact with cellular proteins to carry out at least some its functions. th ... | 1997 | 9350349 |
| recombinant adeno-associated virus type 2 replication and packaging is entirely supported by a herpes simplex virus type 1 amplicon expressing rep and cap. | recombinant adeno-associated virus (aav) type 2 (raav) vectors have recently been shown to have great utility as gene transfer agents both in vitro and in vivo. one of the problems associated with the use of raav vectors has been the difficulty of large-scale vector production. low-efficiency plasmid transfection of the raav vector and complementing aav type 2 (aav-2) functions (rep and cap) followed by superinfection with adenovirus has been the standard approach to raav production. the objecti ... | 1997 | 9343238 |
| adeno-associated virus type 2-mediated transduction in primary human bone marrow-derived cd34+ hematopoietic progenitor cells: donor variation and correlation of transgene expression with cellular differentiation. | although the adeno-associated virus type 2 (aav) is known to possess a broad host range that transcends the species barrier, we suggested in an earlier study that aav infection of human cells is receptor mediated (s. ponnazhagan et al., j. gen. virol. 77:1111-1122, 1996). in the present studies, we investigated the ability of aav to infect primary human hematopoietic progenitor cells capable of multilineage differentiation. bone marrow-derived cd34+ cells from 12 hematologically normal volunteer ... | 1997 | 9343178 |
| high mobility group chromosomal protein 1 binds to the adeno-associated virus replication protein (rep) and promotes rep-mediated site-specific cleavage of dna, atpase activity and transcriptional repression. | high mobility group protein 1 (hmg1) is an abundant non-histone chromosomal protein which plays a role in several nuclear events involving dna. here we demonstrate that hmg1 physically interacts with the human adeno-associated virus (aav) rep protein. hmg1 promotes the formation of rep-dna complexes and stimulates the activity of rep in site- and strand-specific cleavage of dna and the hydrolysis of atp, functions required for viral gene regulation, replication and site-specific integration of v ... | 1997 | 9312052 |
| adeno-associated virus rep proteins target dna sequences to a unique locus in the human genome. | we have developed a system for site-specific dna integration in human cells, mediated by the adeno-associated virus (aav) rep proteins. in its normal lysogenic cycle, aav integrates at a site on human chromosome 19 termed aavs1. we describe a rapid pcr assay for the detection of integration events at aavs1 in whole populations of cells. using this assay, we determined that the aav rep proteins, delivered in cis or trans, are required for integration at aavs1. only the large forms of the rep prot ... | 1997 | 9311886 |
| gene transfer of the costimulatory molecules b7-1 and b7-2 into human multiple myeloma cells by recombinant adeno-associated virus enhances the cytolytic t cell response. | gene transfer of the costimulatory molecules b7-1 and b7-2 induces a potent antitumor immune response in a variety of tumor models. b cell neoplasms including multiple myeloma (mm) often show little or no expression of b7 antigens; they are therefore a potential target for this approach. to increase the expression of human b7 genes in mm cells, both genes and the neomycin phosphotransferase gene were packaged into recombinant adeno-associated virus vectors (raav). the resulting recombinant virus ... | 1997 | 9282174 |
| cancer gene therapy using a novel adeno-associated virus vector expressing human wild-type p53. | previous studies have indicated that transfer of wild-type (wt) p53 cdna into cancer cells can suppress the tumor phenotype in vitro and in vivo. in this study we examined the effects of wt p53 transduction in the human cancer cell line h-358 (that bears a homozygous deletion of p53) using a novel recombinant adeno-associated viral vector engineered to express wt p53 (raavp53). western blot analysis demonstrated the expression of wt p53 in h-358 cells following infection with raavp53. furthermor ... | 1997 | 9282168 |
| cloning of adeno-associated virus type 4 (aav4) and generation of recombinant aav4 particles. | we have cloned and characterized the full-length genome of adeno-associated virus type 4 (aav4). the genome of aav4 is 4,767 nucleotides in length and contains an expanded p5 promoter region compared to aav2 and aav3. within the inverted terminal repeat (itr), several base changes were identified with respect to aav2. however, these changes did not affect the ability of this region to fold into a hairpin structure. within the itr, the terminal resolution site and rep binding sites were conserved ... | 1997 | 9261407 |
| human immunodeficiency virus-1 proviral gene disruption by targeted gene therapy: a hypothetical technique for the elimination of provirus from the infected cells. | a hypothetical technique is proposed for the elimination of all the integrated human immunodeficiency virus-1 provirus from infected cells, based on the developing technology of selective gene excision through homologous recombination. in this technique, a recombinant retroviral packaging cell-line which would produce integrase-rep78 chimeric protein would be constructed. replication defective viral stocks would be made from this system which would have recombinant integrase-rep78 protein packag ... | 1997 | 9247905 |
| infection of primary cells by adeno-associated virus type 2 results in a modulation of cell cycle-regulating proteins. | it has been demonstrated that infection of primary human cells with adeno-associated viruses (aav) leads to a decrease in cellular proliferation and to growth arrest. we analyzed the molecular basis of this phenomenon and observed that infection with aav type 2 (aav2) had an effect on several factors engaged in the control of the mammalian cell cycle. in particular, all of the prb family members, prb, p107, and p130, which are involved in g1 cell cycle checkpoint control, were affected. after in ... | 1997 | 9223493 |
| adeno-associated viral vector gene transfer into leptomeningeal xenografts. | leptomeningeal carcinomatosis is a painful and debilitating complication of cancer. indwelling reservoirs provide continuous assess to the subarachnoid space, making leptomeningeal cancer potentially amenable to gene therapy. adeno-associated virus (aav) is a defective virus not associated with any human disease. we used an aav vector to transduce medulloblastoma (daoy) cells in a nude rat model of leptomeningeal disease. after intraventricular injection of vector carrying the bacterial lacz gen ... | 1997 | 9210060 |
| persistent and therapeutic concentrations of human factor ix in mice after hepatic gene transfer of recombinant aav vectors. | haemophilia b, or factor ix deficiency, is a x-linked recessive disorder that occurs in about one in 25,000 males, and severely affected people are at risk for spontaneous bleeding into numerous organs. bleeding can be life-threatening or lead to chronic disabilities with haemophilic arthropathy. the severity of the bleeding tendency varies among patients and is related to the concentration of functional plasma factor ix. patients with 5-30% of the normal factor ix have mild haemophilia that may ... | 1997 | 9207793 |
| adeno-associated virus (aav) vector antisense gene transfer in vivo decreases gaba(a) alpha1 containing receptors and increases inferior collicular seizure sensitivity. | in the inferior colliculus, adeno-associated virus (aav) vectors are capable of gene transfer and stable, long-term expression, but it remained to be shown if this in vivo gene transfer could alter focal seizure sensitivity in the inferior colliculus. because gaba receptors directly modulate inferior collicular seizures, aav vectors were constructed with a cytomegalovirus (cmv) promoter and a truncated, human gaba(a) alpha1 cdna in both the sense and antisense orientations. seven days after coll ... | 1997 | 9187316 |
| stable gene transfer and expression of human blood coagulation factor ix after intramuscular injection of recombinant adeno-associated virus. | we sought to determine whether intramuscular injection of a recombinant adeno-associated virus (raav) vector expressing human factor ix (hf.ix) could direct expression of therapeutic levels of the transgene in experimental animals. high titer (10(12)-10(13) vector genomes/ml) raav expressing hf.ix was prepared, purified, and injected into hindlimb muscles of c57bl/6 mice and rag 1 mice. in the immunocompetent c57bl/6 mice, immunofluorescence staining of muscle harvested 3 months after injection ... | 1997 | 9159155 |
| gene therapeutic strategies for neuroprotection: implications for parkinson's disease. | gene transfer methodologies are being explored as strategies to restore and preserve neuronal function in parkinson's disease. this technology represents a new therapeutic modality, holding promise for continuous and localized delivery, of neuroprotective molecules. two primary approaches for gene transfer have emerged: in vivo and ex vivo. recent advances in the construction and characterization of gene transfer vectors have generated more efficient vehicles to deliver and express candidate the ... | 1997 | 9126153 |
| detection of adeno-associated virus type 2 in sorted human bone marrow progenitor cells. | wild-type adeno-associated virus (wtaav) is a helper-dependent human parvovirus which has the ability to integrate into the genome of a wide variety of human cells, including those of the hemopoietic lineages. recombinant adeno-associated virus (raav) is becoming a good candidate for virally mediated gene therapy. raav is likely to be a safe vector in clinical gene transfer, as it has never been associated with any disease despite previous studies showing that up to 70% of adults are seropositiv ... | 1997 | 9091303 |
| characterization of recombinant adeno-associated virus-2 as a vehicle for gene delivery and expression into vascular cells. | we have used wild-type and recombinant adeno-associated virus-2 (aav) to study transduction, replication efficiencies, functional protein expression, and gene delivery to vascular cells in vitro and in vivo. | 1997 | 9084579 |
| hsv/aav hybrid amplicon vectors extend transgene expression in human glioma cells. | novel hybrid vectors, which incorporate critical elements of both herpes simplex virus type 1 (hsv-1) amplicon vectors and adeno-associated virus (aav) vectors, are able to sustain transgene expression in dividing glioma cells for over 2 weeks. these vectors combine the high infectibility and large transgene capacity of hsv-1 vectors with the potential for episomal amplification and chromosomal integration of aav vectors. the hybrid vectors contain the hsv-1 origin of dna replication, oris, and ... | 1997 | 9048203 |
| persistent expression of human clotting factor ix from mouse liver after intravenous injection of adeno-associated virus vectors. | we previously found that gene transduction by adeno-associated virus (aav) vectors in cell culture can be stimulated over 100-fold by treatment of the target cells with agents that affect dna metabolism, such as irradiation or topoisomerase inhibitors. here we show that previous gamma-irradiation increased the transduction rate in mouse liver by up to 900-fold, and the topoisomerase inhibitor etoposide increased transduction by about 20-fold. similar rates of hepatic transduction were obtained b ... | 1997 | 9037069 |
| comparison of retroviral and adeno-associated viral vectors designed to express human clotting factor ix. | several different designs for retroviral and adeno-associated virus (aav) vectors were developed to express human clotting factor ix. seven separate retroviral vectors were constructed, including chimeric long terminal repeat (ltr)-based designs, vectors containing splice donor/acceptor sites with internal ribosome entry sites (ires), and vectors with an internal cytomegalovirus (cmv)- or hepatitis b virus (hbv)-derived promoter. five aav vectors were produced using the same cassette design wher ... | 1997 | 9017417 |
| a novel terminal resolution-like site in the adeno-associated virus type 2 genome. | the adeno-associated virus 2 (aav) contains a single-stranded dna genome of which the terminal 145 nucleotides are palindromic and form t-shaped hairpin structures. these inverted terminal repeats (itrs) play an important role in aav dna replication and resolution, since each of the itrs contains a terminal resolution site (trs) that is the target site for the aav rep gene products (rep). however, the rep proteins also interact with the aav dna sequences that lie outside the itrs, and the itrs a ... | 1997 | 8995635 |
| gene delivery to skeletal muscle results in sustained expression and systemic delivery of a therapeutic protein. | somatic gene therapy has been proposed as a means to achieve systemic delivery of therapeutic proteins. however, there is limited evidence that current methods of gene delivery can practically achieve this goal. in this study, we demonstrate that, following a single intramuscular administration of a recombinant adeno-associated virus (raav) vector containing the beta-galactosidase (aav-lacz) gene into adult balb/c mice, protein expression was detected in myofibers for at least 32 weeks. a single ... | 1996 | 8943064 |
| cocrystal structure of yy1 bound to the adeno-associated virus p5 initiator. | ying-yang 1 protein (yy1) supports specific, unidirectional initiation of messenger rna production by rna polymerase ii from two adjacent start sites in the adeno-associated virus p5 promoter, a process which is independent of the tata box-binding protein (tbp). the 2.5-a resolution yy1-initiator element cocrystal structure reveals four zinc fingers recognizing a yy1-binding consensus sequence. upstream of the transcription start sites protein-dna contacts involve both strands and downstream the ... | 1996 | 8942976 |
| a novel adenovirus-adeno-associated virus hybrid vector that displays efficient rescue and delivery of the aav genome. | adenovirus and adeno-associated virus (aav) are eukaryotic dna viruses being developed as vectors for human gene therapy. the strengths of each system have been exploited in a novel vector that is based on an adenovirus-aav hybrid virus incorporated into a plasmid-based molecular conjugate. efficient rescue and replication of the recombinant aav genome in this hybrid required transient expression of rep. this feature was incorporated into the transducing particle by conjugating a rep expression ... | 1996 | 8934222 |
| autonomous parvovirus transduction of a gene under control of tissue-specific or inducible promoters. | several classes of viruses are in use, or are being developed, as gene therapy vectors. viruses with small genomes containing few essential genes have the advantage of requiring only simple complementation systems to allow packaging of foreign dna, substituted for the entire viral coding sequences. retroviruses and the dependent parvovirus aav (adeno-associated virus) have been used in this way, and both possess an efficient integration mechanism which should allow long-term expression of transd ... | 1996 | 8929909 |
| the adeno-associated virus rep78 major regulatory/transformation suppressor protein binds cellular sp1 in vitro and evidence of a biological effect. | adeno-associated virus (aav) rep78 is a multifunctional protein that is required for aav transcriptional activity, aav dna replication, and possibly for site-specific integration of aav into human chromosome 19. rep78 is also able to inhibit a variety of heterologous promoters, including those of c-h-ras, human papillomavirus types 16 and 18, and hiv type 1. however, rep78 is unable to significantly affect murine osteosarcomavirus (msv). it was noticed that promoters that are inhibited possess b ... | 1996 | 8912872 |
| development of adeno-associated virus vectors for gene therapy of cystic fibrosis. | 1996 | 8794249 | |
| gene therapy with an adeno-associated virus carrying an interferon gene results in tumor growth suppression and regression. | adeno-associated virus (aav) vectors were constructed containing both a synthetic type i interferon gene, (ifn-con1) and the bacterial neomycin-resistant gene. recombinant virions were used to infect a number of human tumor cell lines, including 293, hela, k562, and eskol (a hairy cell leukemia-like cell), and geneticin-resistant cells were selected. all ifn-con1-transduced cell lines produced low levels of ifn-con1 and grew at the same rate as nontransduced cell lines. although these cell lines ... | 1996 | 8785709 |
| the recombination signals for adeno-associated virus site-specific integration. | the adeno-associated virus (aav) genome integrates site specifically into a defined region of human chromosome 19 (termed aavs1). using a functional assay for aav integration into aavs1 dna propagated as an episome, we obtained evidence that a 33-nucleotide aavs1 dna sequence contains the minimum signal required for targeted integration. the recombination signal comprises a dna-binding motif for the aav regulatory rep protein [rep binding site (rbs)] separated by an eight-nucleotide spacer from ... | 1996 | 8755586 |
| long-term expression of a fluorescent reporter gene via direct injection of plasmid vector into mouse skeletal muscle: comparison of human creatine kinase and cmv promoter expression levels in vivo. | expression of a fluorescent reporter gene has been studied using two alternate promoters to transcribe the green fluorescent protein (gfp) from aequorea victoria. the human cytomegalovirus (cmv) enhancer/ promoter or the human muscle-specific creatine kinase promoter (ckm) were inserted along with the gfp cdna into a plasmid expression vector based on a modified adeno-associated virus genome. naked plasmid dna was injected into the hamstring muscle of mdx mice and gfp gene expression determined ... | 1996 | 8727010 |
| determination and analysis of the complete nucleotide sequence of human herpesvirus. | human herpesvirus 7 (hhv-7) is a recently isolated betaherpesvirus that is prevalent in the human population, with primary infection usually occurring in early childhood. hhv-7 is related to human herpesvirus 6 (hhv-6) in terms of both biological and, from limited prior dna sequence analysis, genetic criteria. however, extensive analysis of the hhv-7 genome has not been reported, and the precise phylogenetic relationship of hhv-7 to the other human betaherpesviruses hhv-6 and human cytomegalovir ... | 1996 | 8709220 |
| a "humanized" green fluorescent protein cdna adapted for high-level expression in mammalian cells. | we constructed gfph, a synthetic version of the jellyfish aequorea victoria green fluorescent protein (gfp) cdna that is adapted for high-level expression in mammalian cells, especially those of human origin. a total of 92 base substitutions were made in 88 codons in order to change the codon usage within the gfp10 coding sequence so that it was more appropriate for expression in mammalian cells. we also describe a series of versatile recombinant adeno-associated virus and adenovirus vectors for ... | 1996 | 8676491 |
| adeno-associated virus 2-mediated transduction and erythroid cell-specific expression of a human beta-globin gene. | recombinant adeno-associated virus 2 (aav) virions were constructed that contained the genomic copy of a normal human beta-globin gene marked with a 4-bp clal linker, and the herpesvirus thymidine kinase (tk) promoter-driven bacterial gene for resistance to neomycin (v beta m-globin), as well as those containing the dnase l-hypersensitive site 2 (hs-2) from the locus control region (lcr) of the human beta-globin gene cluster (vhs2-beta m-globin). these recombinant virions were used to infect a h ... | 1996 | 8646553 |
| identification of mutant adeno-associated virus rep proteins which are dominant-negative for dna helicase activity. | adeno-associated virus type 2 (aav) rep proteins have been postulated to play a role in unwinding the 145-bp inverted terminal repeats during aav dna replication. previous studies showed that aav rep78 and rep68 could unwind a dna partial duplex of 26 bp. in this work it is demonstrated that nuclear extracts of human 293 cells containing wild-type rep68 can unwind partial dna duplexes up to 160 bp long. mutant rep proteins with either a histidine substituted for lysine 340 or a deletion of methi ... | 1996 | 8645299 |
| recruitment of wild-type and recombinant adeno-associated virus into adenovirus replication centers. | replication of a human parvovirus, adeno-associated virus (aav), is facilitated by coinfection with adeno-virus to provide essential helper functions. we have used the techniques of in situ hybridization and immunocytochemistry to characterize the localization of aav replication within infected cells, previous studies have shown that adenovirus establishes foci called replication centers within the nucleus, where adenoviral replication and transcription occur. our studies indicate that aav is co ... | 1996 | 8627709 |
| parvovirus b19 promoter at map unit 6 confers autonomous replication competence and erythroid specificity to adeno-associated virus 2 in primary human hematopoietic progenitor cells. | the pathogenic human parvovirus b19 is an autonomously replicating virus with a remarkable tropism for human erythroid progenitor cells. although the target cell specificity for b19 infection has been suggested to be mediated by the erythrocyte p-antigen receptor (globoside), a number of nonerythroid cells that express this receptor are nonpermissive for b19 replication. to directly test the role of expression from the b19 promoter at map unit 6 (b19p6) in the erythroid cell specificity of b19, ... | 1995 | 8618912 |
| sero-epidemiological analysis of the risk of virus infections for childhood leukaemia. | virus infections have been thought to be involved in the development of childhood leukaemia. in order to address this issue we determined, in a case-control study, the prevalence of antibodies to viruses infecting blood or bone-marrow cells [epstein-barr virsus (ebv), human herpes virus type 6 (hhv-6), parvovirus b19] as well as to the human virus known for its tumour-suppressive properties, the adeno-associated virus type 2 (aav-2), in the sera of 121 children with leukaemia in germany, and in ... | 1996 | 8598307 |
| adeno-associated virus (aav) as a vector for gene transfer into glial cells of the human central nervous system. | to examine the potential of aav as a vector for gene transfer in glial cells, an established astrocytoma cell line and short-term cultures derived from human oligodendroglioma have been coinfected with aav and helper adenovirus. the level of aav replication in glioma cells was high indicating that they express receptors for aav. | 1994 | 8542426 |
| drug management of noninfective complications of cystic fibrosis. | cystic fibrosis (cf) is the commonest lethal hereditary disease in caucasians. the disease involves a gene mutation located at the long arm of chromosome 7, and more than 300 mutations have been identified. cf is a systemic illness affecting the upper respiratory tract and airways, sweat and salivary glands, pancreas, gastrointestinal tract, liver and male reproductive system. the course is highly variable depending on the specific molecular abnormalities in the mutant gene. the current approach ... | 1995 | 8536551 |
| suppression of human alpha-globin gene expression mediated by the recombinant adeno-associated virus 2-based antisense vectors. | we sought to investigate the usefulness of the adeno-associated virus 2 (aav)-based vectors to suppress the excess production of the human alpha-globin gene product towards developing a treatment modality for beta-thalassemia since accumulation of free alpha-globin reduces the lifespan of red blood cells in these patients. we constructed recombinant aav virions containing the human alpha-globin gene sequences in antisense orientation driven by the herpesvirus thymidine kinase (tk) promoter, the ... | 1994 | 8294880 |
| adeno-associated virus vectors preferentially transduce cells in s phase. | vectors based on adeno-associated virus can stably transfer genes by chromosomal integration in recipient cells. in this study we have infected stationary and dividing primary human fibroblast cultures with adeno-associated virus vectors encoding alkaline phosphatase and neomycin phosphotransferase. we find that the transduction frequency of s phase cells is about 200 times that of non-s phase cells. however, neither s phase nor mitosis is essential for transduction. single-stranded vector genom ... | 1994 | 8090744 |
| adeno-associated virus (aav) rep proteins mediate complex formation between aav dna and its integration site in human dna. | aav is unique among eukaryotic viruses in the ability of its dna to integrate preferentially into a specific region of the human genome. understanding aav integration may aid in developing gene therapy systems with predictable integration sites. using a gel mobility-shift assay, we have identified a dna sequence within the aav integration locus on human chromosome 19 which is specifically bound by the aav rep78 and rep68 proteins. this rep recognition sequence is a gctc repeating motif very simi ... | 1994 | 8016070 |
| adeno-associated virus type 2 interferes with early development of mouse embryos. | the human helper-dependent adeno-associated virus type 2 (aav-2) has been shown to induce differentiation in various cell types in culture including pluripotent embryonic cells, in the absence of helper virus. to assess whether induction of differentiation may influence developmental processes we analysed the effect of aav-2 on developing mouse embryos. in vitro infection of fertilized eggs induced arrest of development at the two-cell stage. moreover, micro injection of aav-2 dna (comprising ei ... | 1994 | 7931151 |
| asymmetric replication in vitro from a human sequence element is dependent on adeno-associated virus rep protein. | the dna of human parvovirus adeno-associated virus type 2 (aav) integrates preferentially into a defined region of human chromosome 19. southern blots of genomic dna from latently infected cell lines revealed that the provirus was not simply inserted into the cellular dna. both the proviral and adjoining cellular dna organization indicated that integration occurred by a complex, coordinated process involving limited dna replication and rearrangements. however, the mechanism for targeted integrat ... | 1995 | 7884849 |
| gene transfer into hematopoietic progenitor and stem cells: progress and problems. | gene transfer to hematopoietic cells for the purpose of "gene therapy" is a new and rapidly developing field with clinical trials in progress. a fundamental goal of research in this field is the incorporation of exogenous genes into the chromosomes of the most primitive hematopoietic progenitor cells--stem cells. recombinantly engineered retroviral vectors are the best characterized and are currently the only vector type in clinical trials directed at the hematopoietic system. high efficiency ge ... | 1994 | 7881358 |
| detection of adeno-associated virus dna in human genital tissue and in material from spontaneous abortion. | the human helper virus-dependent parvovirus, adeno-associated virus (aav) has never been associated with disease in humans [berns et al. (1987): advances in virus research 32:243-306; siegl et al. (1985): intervirology 23:61-73]. however, in pregnant mice, infection with aav induces early abortion [botquin et al. (1993): journal of cancer research and clinical oncology 119:24]. we investigated whether this common human virus may be found in human genital tissue or in curettage material from spon ... | 1994 | 7852963 |
| adenovirus and adeno-associated virus mediated gene transfer. | in this review we describe current strategies for adenoviral mediated gene transfer (amgt) and adeno-associated viral mediated gene transfer (aavmgt). we consider the structure and molecular biology of adenoviruses and adeno-associated viruses and detail the current advantages and disadvantages of amgt and aavmgt. potential solutions to some of the specific drawbacks to amgt, including the development of new vectors, addition of gp19k, organoides, and the use of non-human adenoviral vectors, are ... | 1995 | 7767647 |