| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 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 |
| 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 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| [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 |
| 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 |
| use of the nadh-quinone oxidoreductase (ndi1) gene of saccharomyces cerevisiae as a possible cure for complex i defects in human cells. | the ndi1 enzyme of saccharomyces cerevisiae is a single subunit rotenone-insensitive nadh-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. we have shown previously that the ndi1 gene can be functionally expressed in chinese hamster cells (seo, b. b., kitajima-ihara, t., chan, e. k., scheffler, i. e., matsuno-yagi, a., and yagi, t. (1998) proc. natl. acad. sci. u. s. a. 95, 9167-9171) and human embryonal kidney 293 (hek 293) cells (seo, b. b., matsuno ... | 2000 | 10982813 |
| 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 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 |
| production of recombinant adeno-associated virus. | currently, raav appears to be one of the most promising vectors for gene therapy applications. attractive features of the vector include nonpathogenicity, the ability to infect nondividing cells, escape from host immune responses, and integration into the host genome. tremendous progress has been made in the production of this vector, which makes it possible to start to examine the vector performance in large animals and to implement the transition to phase i human clinical trials with a variety ... | 2000 | 11050955 |
| 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 |
| 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 |
| 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 |
| 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 |
| evolutionary relationships among parvoviruses: virus-host coevolution among autonomous primate parvoviruses and links between adeno-associated and avian parvoviruses. | the current classification of parvoviruses is based on virus host range and helper virus dependence, while little data on evolutionary relationships among viruses are available. we identified and analyzed 472 sequences of parvoviruses, among which there were (virtually) full-length genomes of all 41 viruses currently recognized as individual species within the family parvoviridae. our phylogenetic analysis of full-length genomes as well as open reading frames distinguished three evolutionary gro ... | 2001 | 11222696 |
| 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 |
| 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 |
| gene therapy: a 2001 perspective. | in the past year, three clinical trials of gene therapy for haemophilia have been initiated. years of preclinical studies have culminated in translation of research findings into the clinical arena. it is too early to predict which, if any, of these strategies will show efficacy. this paper will review basic aspects of gene therapy for haemophilia and will briefly outline current clinical trials. the three clinical trials all share a dose escalation design. the ongoing trial for haemophilia b in ... | 2001 | 11240615 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| hybrid vectors based on adeno-associated virus serotypes 2 and 5 for muscle-directed gene transfer. | vectors based on hybrids consisting of adeno-associated virus types 2 (itrs and rep) and 5 (cap) were evaluated for muscle-directed gene transfer (called aav2/5). evaluation in immune-competent mice revealed greater transduction efficacy with aav2/5 than with aav2 and no cross-neutralization between aav2/5 and aav2. interestingly, we saw no immunologic evidence of previous exposure to aav5 capsids in a large population of healthy human subjects. | 2001 | 11390622 |
| adeno-associated virus 2-mediated transduction and erythroid lineage-restricted long-term expression of the human beta-globin gene in hematopoietic cells from homozygous beta-thalassemic mice. | adeno-associated virus 2 (aav), a nonpathogenic human parvovirus, has gained attention as a potentially useful vector for human gene therapy. here, we report successful aav-mediated stable transduction and high-efficiency, long-term, erythroid lineage-restricted expression of a human beta-globin gene in primary murine hematopoietic stem cells in vivo. bone marrow-derived primitive sca-1(+), lin(-) hematopoietic stem cells from homozygous beta-thalassemic mice were transduced ex vivo with a recom ... | 2001 | 11407908 |
| incorporation of tumor-targeting peptides into recombinant adeno-associated virus capsids. | the human parvovirus adeno-associated virus type 2 (aav-2) possesses many features that make it an attractive vector for gene delivery in vivo. however, its broad host range may limit its usefulness and effectivity in several gene therapy applications in which transgene expression needs to be limited to a specific organ or cell type. in this study, we explored the possibility of directing recombinant aav-2 transduction by incorporating targeting peptides previously isolated by in vivo phage disp ... | 2001 | 11407911 |
| adeno-associated virus 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 |
| 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 |
| design and packaging of adeno-associated virus gene targeting vectors. | adeno-associated virus (aav) vectors can transduce cells by several mechanisms, including (i) gene addition by chromosomal integration or episomal transgene expression or (ii) gene targeting by modification of homologous chromosomal sequences. the latter process can be used to correct a variety of mutations in chromosomal genes with high fidelity and specificity. in this study, we used retroviral vectors to introduce mutant alkaline phosphatase reporter genes into normal human cells and subseque ... | 2000 | 10775597 |
| gene delivery to human chondrocytes by an adeno associated virus vector. | to investigate the efficiency of gene transduction to human chondrocytes using an adeno associated virus (aav) vector. | 2000 | 10782826 |
| increasing the size of raav-mediated expression cassettes in vivo by intermolecular joining of two complementary vectors. | a major shortcoming to the use of adeno-associated virus (raav) vectors is their limited packaging size. to overcome this hurdle, we split an expression cassette and cloned it into two separate vectors. the vectors contained either a nuclear localizing escherichia coli lacz transgene (nlslacz) with a splice acceptor, or the human elongation factor 1alpha ( ef1alpha) gene enhancer/promoter(s) (ef1alphaep) with a splice donor. we co-injected a promoter-less nlslacz vector with a vector containing ... | 2000 | 10802620 |
| aspartoacylase gene transfer to the mammalian central nervous system with therapeutic implications for canavan disease. | with the ultimate goal of developing safe and effective in vivo gene therapy for the treatment of canavan disease and other neurological disorders, we developed a non-viral lipid-entrapped, polycation-condensed delivery system (lpd) for central nervous system gene transfer, in conjunction with adeno-associated virus (aav)-based plasmids containing recombinant aspartoacylase (aspa). the gene delivery system was tested in healthy rodents and primates, before proceeding to preliminary studies in 2 ... | 2000 | 10894213 |
| recombinant adeno-associated virus vectors for cystic fibrosis gene therapy. | cystic fibrosis (cf) is an autosomal recessive inherited disorder that affects approximately 30,000 north americans. defects in the cf transmembrane conductance regulator (cftr) gene lead to altered secretions from exocrine glands and the pulmonary airways, to a heightened susceptibility to airway infections with pseudomonas aeruginosa, and to severe airway inflammation. early attempts to develop a genetic therapy for cf have not met with great clinical success, but these efforts have driven the ... | 2001 | 11699895 |
| inhibition of s-phase progression by adeno-associated virus rep78 protein is mediated by hypophosphorylated prb. | adeno-associated virus (aav) has an antiproliferative action on cells. we investigated the effect of the aav replication proteins (rep) on the cell division cycle using retroviral vectors. rep78 and rep68 inhibited the growth of primary, immortalized and transformed cells, while rep52 and rep40 did not. rep68 induced cell cycle arrest in phases g(1) and g(2), with elevated cdk inhibitor p21 and reduced cyclin e-, a- and b1-associated kinase activity. rep78-expressing cells were also impaired in ... | 2000 | 10944118 |
| gene therapy for hypertension: sense and antisense strategies. | gene therapy for hypertension is needed for the next generation of antihypertensive drugs. current drugs, although effective, have poor compliance, are expensive and short-lasting (hours or one day). gene therapy offers a way to produce long-lasting antihypertensive effects (weeks, months or years). we are currently using two strategies: antisense oligodeoxynucleotides (as-odn), an dantisense dna delivered in viral vectors, to inhibit genes associated with vasoconstrictive properties. it is not ... | 2001 | 11727501 |
| delivery of novel macromolecular drugs against hiv-1. | the development of new low molecular weight drugs against human immunodeficiency virus type 1 (hiv-1) targets other than reverse transcriptase (rt) and protease, such as the integrase and the envelope glycoprotein, is likely to take many years. macromolecular drugs, including antisense oligonucleotides, ribozymes, rna decoys and transdominant mutant proteins, may be able to interfere with a relatively large number of viral targets, thereby decreasing the likelihood of the emergence of drug-resis ... | 2001 | 11728227 |
| hyaluronidase enhances recombinant adeno-associated virus (raav)-mediated gene transfer in the rat skeletal muscle. | skeletal muscle is a privileged target for long-term raav-mediated gene transfer in mouse, rat, dog and non-human primates. intramuscular injections of raav encoding human factor ix in hemophilia b patients have been initiated, based on promising results gathered in affected dogs. we found that intramuscular raav administration in rats resulted in restricted transduction essentially along the myofibers axis with poor lateral diffusion. this suggested that the transduction rate might be limited b ... | 2000 | 10981669 |
| exchange of surface proteins impacts on viral vector cellular specificity and transduction characteristics: the retina as a model. | recombinant vectors based on adeno-associated virus (aav) or human immunodeficiency 1 (lentivirus) are promising tools for long term in vivo gene delivery. their design allows the exchange of capsids or envelopes, respectively, theoretically providing the opportunity to transduce a range of cell types. we constructed aav vectors encoding enhanced green fluorescent protein (egfp) within an aav serotype 2 (aav2) genome contained in an aav2, five or one capsid (called aav2/2, aav2/5 and aav2/1, res ... | 2001 | 11751689 |
| a high-capacity, capsid-modified hybrid adenovirus/adeno-associated virus vector for stable transduction of human hematopoietic cells. | to achieve stable gene transfer into human hematopoietic cells, we constructed a new vector, deltaad5/35.aav. this vector has a chimeric capsid containing adenovirus type 35 fibers, which conferred efficient infection of human hematopoietic cells. the deltaad5/35.aav vector genome is deleted for all viral genes, allowing for infection without virus-associated toxicity. to generate high-capacity deltaad5/35.aav vectors, we employed a new technique based on recombination between two first-generati ... | 2002 | 11773389 |
| selective cleavage of aavs1 substrates by the adeno-associated virus type 2 rep68 protein is dependent on topological and sequence constraints. | the adeno-associated virus type 2 (aav-2) rep78 and rep68 proteins are required for replication of the virus as well as its site-specific integration into a unique site, called aavs1, of human chromosome 19. rep78 and rep68 initiate replication by binding to a rep binding site (rbs) contained in the aav-2 inverted terminal repeats (itrs) and then specifically nicking at a nearby site called the terminal resolution site (trs). similarly, rep78 and rep68 are postulated to trigger the integration p ... | 2000 | 10982325 |
| adeno-associated virus type 2 rep78 induces apoptosis through caspase activation independently of p53. | adeno-associated virus (aav) type 2 rep78 is a multifunctional protein required for aav dna replication, integration, and gene regulation. the biochemical activities of rep78 have been described, but the effects of rep proteins on the cell have not been characterized. we have analyzed rep-mediated cytotoxicity. we demonstrated that rep78 expression is sufficient to induce cell death and disruption of the cell cycle. cell death was found to be mediated by apoptosis. rep78 expression resulted in t ... | 2000 | 11000213 |
| adeno-associated virus vector mediated gene transfer to pancreatic beta cells. | insulin-dependent diabetes mellitus (iddm) or type 1 diabetes is an autoimmune disease that results in destruction of the insulin-producing pancreatic islet beta cells. several factors induce the invasion of immune cells into islets and trigger inflammation. gene therapy approaches targeting the islet cells could be an effective treatment to prevent the onset or reverse type 1 diabetes. allogeneic islet transplantation provides short-term treatment. however, genetically modified islets, which re ... | 2000 | 11021593 |
| recombinant adeno-associated virus serotype 2 vectors mediate stable interleukin 10 secretion from salivary glands into the bloodstream. | we have constructed a recombinant adeno-associated virus serotype 2 vector encoding human interleukin 10 (raavhil10). il-10 is a potent antiinflammatory/immune cytokine, which has received growing attention for its therapeutic potential. human il-10 (hil-10) production was virus dose dependent after in vitro infection of hsg cells, a human submandibular gland cell line. the vector-derived hil-10 produced was biologically active, as the medium from raavhil10-infected hsg cells caused a dose-depen ... | 2002 | 11812284 |
| adeno-associated virus vector carrying human minidystrophin genes effectively ameliorates muscular dystrophy in mdx mouse model. | duchenne muscular dystrophy (dmd) is the most common and lethal genetic muscle disorder, caused by recessive mutations in the dystrophin gene. one of every 3,500 males suffers from dmd, yet no treatment is currently available. genetic therapeutic approaches, using primarily myoblast transplantation and adenovirus-mediated gene transfer, have met with limited success. adeno-associated virus (aav) vectors, although proven superior for muscle gene transfer, are too small (5 kb) to package the 14-kb ... | 2000 | 11095710 |
| adeno-associated virus production of soluble tumor necrosis factor receptor neutralizes tumor necrosis factor alpha and reduces arthritis. | the major limitation of adenovirus is its association with induction of an inflammatory response and relatively short-term production of the gene therapy transgene product. adeno-associated virus (aav) is a 4.68-kb single-strand dna virus that contains itrs for viral replication and a packaging signal, and also has been engineered to contain therapeutic genes up to 5 kb in length. transduction of recombinant aav (raav) results in low inflammatory response and long-term expression. we have cloned ... | 2000 | 11096446 |
| intracellular trafficking of adeno-associated virus vectors: routing to the late endosomal compartment and proteasome degradation. | the early steps of adeno-associated virus (aav) infection involve attachment to a variety of cell surface receptors (heparan sulfate, integrins, and fibroblast growth factor receptor 1) followed by clathrin-dependent or independent internalization. here we have studied the subsequent intracellular trafficking of aav particles from the endosomal compartment to the nucleus. human cell lines were transduced with a recombinant aav (raav) carrying a reporter gene (luciferase or green fluorescent prot ... | 2001 | 11160681 |
| isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column. | one of the most promising gene transfer vectors in human clinical trials is aav2. the quality of the vector preparations is a key element in obtaining reliable and reproducible data in preclinical studies. however, established protocols either result in impure, low infectious virus (cscl2 gradient centrifugation) or demand a high level of manual and technical skills (cscl2 gradient centrifugation, iodixanol/heparin or hplc purification). in this study, we present an easy-to-do single-step column ... | 2001 | 11177544 |
| 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 |
| 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 |
| effect of dna-dependent protein kinase on the molecular fate of the raav2 genome in skeletal muscle. | we report here that the dna-dependent protein kinase (dna-pk) affects the molecular fate of the recombinant adeno-associated virus (raav) genome in skeletal muscle. raav-human alpha1-antitrypsin (raav-haat) vectors were delivered by intramuscular injection to either c57bl/6 (dna-pkcs(+)) or c57bl/6-scid [severe combined immunodeficient (scid), dna-pkcs(-)] mice. in both strains, high levels of transgene expression were sustained for up to 1 year after a single injection. southern blot analysis s ... | 2001 | 11274433 |
| regulated secretion of proinsulin/insulin from human hepatoma cells transduced by recombinant adeno-associated virus. | to employ hepatocytes as surrogate beta-cells for gene therapy of diabetes, a regulatory system was devised in this study by placing the human insulin cdna under the control of the phosphoenolpyruvate carboxykinase (pepck) promoter, followed by the cytomegalovirus immediate early promoter-driven enhanced-green-fluorescent-protein open reading frame. the expression cassette was inserted into the adeno-associated virus vector between two inverted terminal repeats, and used to produce recombinant a ... | 2001 | 11277867 |
| suicide gene therapy for human oral squamous cell carcinoma cell lines with adeno-associated virus vector. | the purpose of this study was to test the possibility of gene transfer as a new therapy for oral cancer. adeno-associated virus (aav) has already been used in the fields of cystic fibrosis and parkinson's disease as a potential vector for gene therapy because of its wide host range, high transduction efficiency, and lack of cytopathogenicity. four human oral squamous cell carcinoma cell lines were transduced with an aav vector containing the beta-galactosidase gene (aavlacz) in vitro. gene trans ... | 2001 | 11287273 |
| adeno-associated virus (aav)-mediated gene transfer in respiratory epithelium and submucosal gland cells in human fetal tracheal organ culture. | since the discovery of the cystic fibrosis transmembrane regulator (cftr) gene, cystic fibrosis has been an attractive target for gene therapy. postnatal gene transfer in the respiratory epithelium has been difficult and particularly inefficient in the submucosal gland cells, the target cells for cftr gene transfer. the authors hypothesized that during development, there is a favorable environment for fetal gene therapy with fewer physical barriers to efficient gene transfer and more accessible ... | 2002 | 12077770 |
| gene therapy restores vision in a canine model of childhood blindness. | the relationship between the neurosensory photoreceptors and the adjacent retinal pigment epithelium (rpe) controls not only normal retinal function, but also the pathogenesis of hereditary retinal degenerations. the molecular bases for both primary photoreceptor and rpe diseases that cause blindness have been identified. gene therapy has been used successfully to slow degeneration in rodent models of primary photoreceptor diseases, but efficacy of gene therapy directed at photoreceptors and rpe ... | 2001 | 11326284 |
| enhancement of muscle gene delivery with pseudotyped adeno-associated virus type 5 correlates with myoblast differentiation. | adeno-associated virus (aav)-based muscle gene therapy has achieved tremendous success in numerous animal models of human diseases. recent clinical trials with this vector have also demonstrated great promise. however, to achieve therapeutic benefit in patients, large inocula of virus will likely be necessary to establish the required level of transgene expression. for these reasons, efforts aimed at increasing the efficacy of aav-mediated gene delivery to muscle have the potential for improving ... | 2001 | 11462038 |
| second generation adeno-associated virus type 2-based gene therapy systems with the potential for preferential integration into aavs1. | adeno-associated virus type 2 (aav-2) is a non-pathogenic human parvovirus that is being developed as a gene therapy vector for the treatment of numerous diseases. one property of wild-type aav-2, that is highly desirable in a gene therapy vector, is its ability to preferentially integrate its dna into a 4 kilobase region of human chromosome 19, designated aavs1. one disadvantage of aav-2 is its relatively small packaging capacity, approximately 4.7 kilobases. because of this size limitation, th ... | 2002 | 12109212 |
| recombinant human parvovirus b19 vectors. | in an attempt to exploit the remarkable tissue-tropism of the human parvovirus b19 to target human hematopoietic cells of the erythroid lineage, recombinant human adeno-associated virus 2 genomes were encapsidated in parvovirus b19 capsids. although efficient transduction of primary human hematopoietic cells in the erythroid lineage occurred, a low-level of transgene expression in non-erythroid cells was also detected. these studies suggest that cell surface expression of p antigen, the primary ... | 2002 | 12116848 |
| characterization of permanent cell lines that contain the aav2 rep-cap genes on an epstein-barr-virus-based episomal plasmid. | recombinant adeno-associated virus (raav) has emerged as a promising gene therapy vector. its development, however, has been hampered by the lack of a readily available efficient production method. we investigated the possibility of establishing permanent cell lines for the production of raav with a new epstein-barr-virus (ebv)-based episomal aav rep-cap plasmid (pcep-rep/cap). hela and 293 cells were stably transfected with plasmids that carry the aav2 rep/cap genes under transcriptional contro ... | 2001 | 11509889 |
| adeno-associated virus type 2-mediated gene transfer: role of cellular fkbp52 protein in transgene expression. | although adeno-associated virus type 2 (aav) has gained attention as a potentially useful vector for human gene therapy, the transduction efficiencies of aav vectors vary greatly in different cells and tissues in vitro and in vivo. we have documented that a cellular tyrosine phosphoprotein, designated the single-stranded d-sequence-binding protein (ssd-bp), plays a crucial role in aav-mediated transgene expression (k. y. qing, x.-s. wang, d. m. kube, s. ponnazhagan, a. bajpai, and a. srivastava, ... | 2001 | 11533160 |
| the atomic structure of adeno-associated virus (aav-2), a vector for human gene therapy. | the structure of the adeno-associated virus (aav-2) has been determined to 3-a resolution by x-ray crystallography. aav is being developed as a vector for gene therapy to treat diseases including hemophilia, cancer, and cystic fibrosis. as in the distantly related autonomous parvoviruses, the capsid protein has a beta-barrel fold, but long loops between the beta-strands share little structural homology with other parvoviruses, leading to unique surface features. most prominent are groups of thre ... | 2002 | 12136130 |
| insertional mutagenesis of the adeno-associated virus type 2 (aav2) capsid gene and generation of aav2 vectors targeted to alternative cell-surface receptors. | recombinant adeno-associated virus (aav) vectors are of interest in the context of gene therapy because of their ability to mediate efficient transfer and stable expression of therapeutic genes in a wide variety of tissues. however, aav-mediated gene delivery to specific cell populations is often precluded by the widespread distribution of heparan sulfate proteoglycan (hspg), the primary cellular receptor for the virus. conversely, an increasing number of cell types are being identified that do ... | 2001 | 11560765 |
| stable therapeutic serum levels of human alpha-1 antitrypsin (aat) after portal vein injection of recombinant adeno-associated virus (raav) vectors. | previous work from our group showed that recombinant adeno-associated virus (raav) vectors mediated long-term secretion of therapeutic serum levels of human alpha-1 antitrypsin (haat) after a single injection in murine muscle. we hypothesized that hepatocyte transduction could be even more efficient, since these cells represent the natural site of aat production and secretion. to test this hypothesis, raav vectors containing the haat cdna driven by either the human elongation factor 1 alpha prom ... | 2001 | 11571566 |
| adeno-associated virus-mediated delivery of glial cell line-derived neurotrophic factor protects motor neuron-like cells from apoptosis. | motor neuron disorders including amyotrophic lateral sclerosis may benefit from the induction of neurotrophic factors such as glial cell line-derived neurotrophic factor (gdnf) that are known to be trophic and protective for motor neurons. however, the application of such factors is limited by an inability to successfully target their expression in the nervous system. in this study we investigate the potential of using adeno-associated virus (aav) as a vector for gene delivery into motor neuron- ... | 2001 | 11582516 |
| adeno-associated virus vector-mediated il-10 gene delivery prevents type 1 diabetes in nod mice. | the development of spontaneous autoimmune diabetes in nonobese diabetic (nod) mice provides for their use as a model of human type 1 diabetes. to test the feasibility of muscle-directed gene therapy to prevent type 1 diabetes, we developed recombinant adeno-associated virus (raav) vectors containing murine cdnas for immunomodulatory cytokines il-4 or il-10. skeletal muscle transduction of female nod mice with il-10, but not il-4, completely abrogated diabetes. raav-il-10 transduction attenuated ... | 2001 | 11717448 |
| glial cell line derived neurotrophic factor delays photoreceptor degeneration in a transgenic rat model of retinitis pigmentosa. | we designed experiments to evaluate the therapeutic potential of glial cell line derived neurotrophic factor (gdnf) to rescue photoreceptors from genetically determined cell death. gene transfer of the neurotrophic factor to the retina was achieved via a recombinant adeno-associated virus (raav) vector containing the chicken beta-actin promoter/immediate early cytomegalovirus enhancer (cba) driving the human gdnf gene. we delivered aav-cba-gdnf to the retinas of an animal model of retinitis pigm ... | 2001 | 11735347 |
| rapid induction of cytotoxic t-cell response against cervical cancer cells by human papillomavirus type 16 e6 antigen gene delivery into human dendritic cells by an adeno-associated virus vector. | we have shown that the pulsing of dendritic cells (dcs) with human papillomavirus type 16 (hpv-16) antigen proteins by lipofection stimulates class i-restricted cytotoxic t lymphocyte (ctl) response against primary cervical cancer cells. also, we have shown that adeno-associated virus (aav) was able to effectively deliver a cytokine gene into dcs. it has been our hypothesis that the delivery of antigen genes into dcs, resulting in endogenous and continuous antigen protein expression, may result ... | 2001 | 11781657 |
| neurological correction of lysosomal storage in a mucopolysaccharidosis iiib mouse model by adeno-associated virus-mediated gene delivery. | mucopolysaccharidosis (mps) iiib is characterized by mild somatic features and severe neurological diseases leading to premature death. no definite treatment is available for mps iiib patients. we constructed two recombinant adeno-associated virus (raav) vectors containing the human alpha-n-acetylglucosaminidase (naglu) cdna driven by either a cmv or a neuron-specific enolase (nse) promoter. in vitro, these raav vectors mediated efficient expression of recombinant naglu in human mps iiib fibrobl ... | 2002 | 11786044 |
| rescue of hereditary form of dilated cardiomyopathy by raav-mediated somatic gene therapy: amelioration of morphological findings, sarcolemmal permeability, cardiac performances, and the prognosis of to-2 hamsters. | the hereditary form comprises approximately 1/5 of patients with dilated cardiomyopathy (dcm) and is a major cause of advanced heart failure. medical and socioeconomic settings require novel treatments other than cardiac transplantation. to-2 strain hamsters with congenital dcm show similar clinical and genetic backgrounds to human cases that have defects in the delta-sarcoglycan (delta-sg) gene. to examine the long-term in vivo supplement of normal delta-sg gene driven by cytomegalovirus promot ... | 2002 | 11805334 |
| [somatic gene therapy of dilated cardiomyopathy]. | the hereditary form of dilated cardiomyopathy (dcm) accounts for about 20% of human dcm and is a major cause of heart failure. to-2 strain hamsters show dcm, a gene deletion of delta-sarcoglycan (sg), loss of all four sgs, alpha-, beta-, gamma- and delta-sg proteins, and are useful for developing gene therapy of the hereditary dcm. the delta-sg is a component of dystrophin-associated glycoprotein complex that stabilizes sarcolemma. four familial and sporadic dcm cases have been reported in human ... | 2002 | 11862755 |
| recombinant adeno-associated virus gene therapy for cystic fibrosis and alpha(1)-antitrypsin deficiency. | | 2002 | 11893723 |
| aav-mediated delivery of ciliary neurotrophic factor prolongs photoreceptor survival in the rhodopsin knockout mouse. | retinitis pigmentosa (rp), an inherited retinal degenerative disease causing blindness, is characterized by progressive apoptotic death of photoreceptors. therapeutic modification of photoreceptor apoptosis may provide an effective therapy for this disorder. ciliary neurotrophic factor (cntf) has been shown to promote survival of a number of different neuronal cell types, including photoreceptors. the present study aimed to test whether adeno-associated virus (aav)-mediated delivery of the gene ... | 2001 | 11237681 |
| epitope-tagged recombinant aav vectors for expressing neurturin and its receptor in retinal cells. | neurturin (ntn) is a potent neuronal survival factor in the central and peripheral nervous systems. we previously described altered expression of mrnas for ntn and one of its receptor components, gfra-2 in degenerative retinas of rd/rd mice. towards assessing the potential for transfer of these genes to counteract retinal degeneration, we examined recombinant adeno-associated virus (raav) constructs for expression of ntn and gfra-2 transgenes in retinal cells in vitro and for the effect of trans ... | 2001 | 11239244 |
| efficient ex vivo transduction of pancreatic islet cells with recombinant adeno-associated virus vectors. | the ability to transfer immunoregulatory, cytoprotective, or antiapoptotic genes into pancreatic islet cells may allow enhanced posttransplantation survival of islet allografts and inhibition of recurrent autoimmune destruction of these cells in type 1 diabetes. however, transient transgene expression and the tendency to induce host inflammatory responses have limited previous gene delivery studies using viral transfer vectors. we demonstrate here that recombinant adeno-associated virus (raav) s ... | 2001 | 11246870 |
| expression of human factor viii by splicing between dimerized aav vectors. | adeno-associated virus (aav) is a useful vector for hemophilia gene therapy, but the limited effective packaging capacity of aav (5 kb) appears to be incompatible with factor viii (gene symbol f8) cdna (7 kb). although we previously demonstrated efficient packaging and expression of b-domain-deleted human f8 (bdd-f8) using a single aav vector, the packaging limit still excludes the use of large/strong regulatory elements. here we exploited the split aav vector technology that expands the packagi ... | 2002 | 12027555 |
| analysis of adeno-associated virus-mediated ex vivo transferred human beta-globin gene in bone marrow engrafted mice. | adeno-associated virus (aav)-2 was developed as a useful vector for human gene therapy. in this report, we analyzed the integration and expression of aav-mediated ex vivo transferred human beta-globin gene in bone marrow (bm) reconstituted mice. recombinant aav (raav) containing human beta-globin gene was packaged by infecting individual g418-resistant bhk-21 cell clones integrated with the plasmid av53hs432deltabeta2.0neo with recombinant herpes simplex virus, which can express rep and cap gene ... | 2002 | 12065900 |
| targeted transgene insertion into human chromosomes by adeno-associated virus vectors. | efficient methods are needed for the precise genetic manipulation of diploid human cells, in which cellular senescence and low conventional gene targeting rates limit experimental and therapeutic options. we have shown previously that linear, single-stranded dna vectors based on adeno-associated virus (aav) could accurately introduce small (<20 bp) genetic modifications into homologous human chromosomal sequences. here we have used aav vectors to introduce large (>1 kb) functional transgene cass ... | 2002 | 12089561 |