Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
|---|
| 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 |
| 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 viral-mediated catalase expression suppresses optic neuritis in experimental allergic encephalomyelitis. | suppression of oxidative injury by viral-mediated transfer of the human catalase gene was tested in the optic nerves of animals with experimental allergic encephalomyelitis (eae). eae is an inflammatory autoimmune disorder of primary central nervous system demyelination that has been frequently used as an animal model for the human disease multiple sclerosis (ms). the optic nerve is a frequent site of involvement common to both eae and ms. recombinant adeno-associated virus containing the human ... | 1998 | 9811889 |
| adeno-associated virus vectors can be efficiently produced without helper virus. | the purpose of this work was to develop an efficient method for the production of adeno-associated virus (aav) vectors in the absence of helper virus. the adenovirus regions that mediate aav vector replication were identified and assembled into a helper plasmid. these included the va, e2a and e4 regions. when this helper plasmid was cotransfected into 293 cells, along with plasmids encoding the aav vector, and rep and cap genes, aav vector was produced as efficiently as when using adenovirus inf ... | 1998 | 9813665 |
| gene modification of primary tumor cells for active immunotherapy of human breast and ovarian cancer. | we have previously shown that cationic liposomes facilitate adeno-associated virus (aav) plasmid transfections of primary and cultured cell types. to test the clinical feasibility of using genetically modified tumor vaccines for the treatment of breast and ovarian cancers, we have constructed an expression plasmid pmp6il2 and investigated the use of liposome-mediated gene delivery into primary, uncultured human breast and ovarian tumor cells to produce interleukin 2 (il-2)-secreting tumor cells. ... | 1996 | 9816091 |
| 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 |
| recent advances in gene therapy of gi and liver diseases. | gene therapy toward the digestive organs has made substantial progress. the strategies applied include ex vivo and in vivo delivery. the ex vivo pathway does not require a tissue-specific vector while the in vivo pathway is advantageous for tissues not obtainable or hard to culture. the specific vectors of gene delivery for the liver and gastrointestinal tract include (1) viral vectors: retrovirus, adenovirus, and adeno-associated virus are the three most common currently used. (2) liposomes: wh ... | 1998 | 9823673 |
| sustained secretion of human alpha-1-antitrypsin from murine muscle transduced with adeno-associated virus vectors. | recombinant adeno-associated virus (aav) vectors have been used to transduce murine skeletal muscle as a platform for secretion of therapeutic proteins. the utility of this approach for treating alpha-1-antitrypsin (aat) deficiency was tested in murine myocytes in vitro and in vivo. aav vectors expressing the human aat gene from either the cytomegalovirus (cmv) promoter (aav-c-at) or the human elongation factor 1-alpha promoter (aav-e-at) were examined. in vitro in c2c12 murine myoblasts, the ex ... | 1998 | 9826709 |
| high-titer adeno-associated viral vectors from a rep/cap cell line and hybrid shuttle virus. | adeno-associated virus (aav) is a potential vector for in vivo gene therapy. a critical analysis of its utility has been hampered by methods of production that are inefficient, difficult to scale up, and that often generate substantial quantities of replication-competent aav. we describe a novel method for producing aav that addresses these problems. a cell line, called b50, was created by stably transfecting into hela cells a rep/cap-containing plasmid utilizing endogenous aav promoters. produc ... | 1998 | 9829534 |
| gene transfer: a review of methods and applications. | gene transfer is a potentially powerful tool for the treatment of a wide variety of diseases. the transfer of these genes is achieved by utilizing a variety of vectors, including retroviral, adenoviral, adeno-associated virus (aav) and a number of non-viral mechanisms. numerous studies have successfully demonstrated transduction of genes into target cells with a variety of vectors, and have provided 'proof-in-principle' that gene transfer can result in prolonged in vivo expression of transduced ... | 1998 | 9839307 |
| developing vdept for dt-diaphorase (nqo1) using an aav vector plasmid. | one enzyme/prodrug combination that has the potential to be used in virally directed enzyme/prodrug therapy (vdept) is the obligate 2-electron reducing enzyme, dt-diaphorase (nqo1), with bioreductive agents such as eo9. the present studies were undertaken to determine if this enzyme, as well as the reporter molecule, green fluorescent protein (gfp), could be expressed from a single dicistronic unit under control of the cmv promoter in an adeno-associated virus (aav) background. | 1998 | 9845120 |
| formation of adeno-associated virus circular genomes is differentially regulated by adenovirus e4 orf6 and e2a gene expression. | a central feature of the adeno-associated virus (aav) latent life cycle is persistence in the form of both integrated and episomal genomes. however, the molecular processes associated with episomal long-term persistence of aav genomes are only poorly understood. to investigate these mechanisms, we have utilized a recombinant aav (raav) shuttle vector to identify circular aav intermediates from transduced hela cells and primary fibroblasts. the unique structural features exhibited by these transd ... | 1999 | 9847318 |
| the adeno-associated virus type 2 regulatory proteins rep78 and rep68 interact with the transcriptional coactivator pc4. | the adeno-associated virus type 2 (aav-2) rep78/rep68 regulatory proteins are pleiotropic effectors of viral and cellular dna replication, of cellular transformation by viral and cellular oncogenes, and of homologous and heterologous gene expression. to search for cellular proteins involved in mediating these functions, we used rep68 as bait in the yeast two-hybrid system and identified the transcriptional coactivator pc4 as a rep interaction partner. pc4 has been shown to mediate transcriptiona ... | 1999 | 9847329 |
| behavioral recovery in 6-hydroxydopamine-lesioned rats by cotransduction of striatum with tyrosine hydroxylase and aromatic l-amino acid decarboxylase genes using two separate adeno-associated virus vectors. | parkinson's disease (pd) is characterized by the progressive loss of the dopaminergic neurons in the substantia nigra and a severe decrease in dopamine in the striatum. a promising approach to the gene therapy of pd is intrastriatal expression of enzymes in the biosynthetic pathway for dopamine. tyrosine hydroxylase (th) catalyzes the synthesis of l-dopa, which must be converted to dopamine by aromatic l-amino acid decarboxylase (aadc). since the endogenous aadc activity in the striatum is consi ... | 1998 | 9853519 |
| 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-mediated delivery of antiangiogenic factors as an antitumor strategy. | antiangiogenic tumor therapies have recently attracted intense interest for their broad-spectrum action, low toxicity, and, in the case of direct endothelial targeting, an absence of drug resistance. to promote tumor regression and to maintain dormancy, antiangiogenic agents need to be chronically administered. gene therapy offers a potential way to achieve sustained therapeutic release of potent antiangiogenic substances. as a step toward this goal, we have generated recombinant adeno-associate ... | 1998 | 9865720 |
| regulated delivery of therapeutic proteins after in vivo somatic cell gene transfer. | stable delivery of a therapeutic protein under pharmacologic control was achieved through in vivo somatic gene transfer. this system was based on the expression of two chimeric, human-derived proteins that were reconstituted by rapamycin into a transcription factor complex. a mixture of two adeno-associated virus vectors, one expressing the transcription factor chimeras and one containing erythropoietin (epo) under the control of a promoter responsive to the transcription factor, was injected in ... | 1999 | 9872748 |
| 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 |
| human adeno-associated virus type 5 is only distantly related to other known primate helper-dependent parvoviruses. | we have characterized 95% (4,404 nucleotides) of the genome of adeno-associated virus type 5 (aav5), including part of the terminal repeats and the terminal resolution site. our results show that aav5 is different from all other described aav serotypes at the nucleotide level and at the amino acid level. the sequence homology to aav2, aav3b, aav4, and aav6 at the nucleotide level is only between 54 and 56%. the positive strand contains two large open reading frames (orfs). the left orf encodes t ... | 1999 | 9882294 |
| cloning and characterization of adeno-associated virus type 5. | adeno-associated virus type 5 (aav5) is distinct from other dependovirus serotypes based on dna hybridization and serological data. to better understand the biology of aav5, we have cloned and sequenced its genome and generated recombinant aav5 particles. the single-stranded dna genome is similar in length and genetic organization to that of aav2. the rep gene of aav5 is 67% homologous to aav2, with the majority of the changes occurring in the carboxyl and amino termini. this homology is much le ... | 1999 | 9882336 |
| biochemical characterization of adeno-associated virus rep68 dna helicase and atpase activities. | the adeno-associated virus (aav) nonstructural proteins rep68 and rep78 are site-specific dna binding proteins, atp-dependent site-specific endonucleases, helicases, and atpases. these biochemical activities are required for viral dna replication and control of viral gene expression. in this study, we characterized the biochemical properties of the helicase and atpase activities of homogeneously pure rep68. the enzyme exists as a monomer in solution at the concentrations used in this study (<380 ... | 1999 | 9882364 |
| human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2. | adeno-associated virus 2 (aav)-based vectors have gained attention as a potentially useful alternative to the more commonly used retroviral and adenoviral vectors for human gene therapy. although aav uses the ubiquitously expressed cell surface heparan sulfate proteoglycan (hspg) as a receptor, the transduction efficiency of aav vectors varies greatly in different cells and tissues in vitro and in vivo. we demonstrate here that cell surface expression of hspg alone is insufficient for aav infect ... | 1999 | 9883842 |
| alphavbeta5 integrin: a co-receptor for adeno-associated virus type 2 infection. | understanding the primary steps of viral entry can have important implications for strategies to prevent infection of known viral pathogens as well as determining parameters for efficient gene delivery using viral vectors. recently, a two-step process for viral infection involving attachment of virus to a primary receptor (coxsackievirus adenovirus receptor and heparan sulfate proteoglycan) and subsequent mediation of virus entry by a co-receptor (alphav integrins and hvem) has been determined f ... | 1999 | 9883843 |
| efficient and stable transduction of cardiomyocytes after intramyocardial injection or intracoronary perfusion with recombinant adeno-associated virus vectors. | the delivery of recombinant genes to cardiomyocytes holds promise for the treatment of a variety of cardiovascular diseases. previous gene transfer approaches that used direct injection of plasmid dna or replication-defective adenovirus vectors have been limited by low transduction frequencies and transient transgene expression due to immune responses, respectively. in this report, we have tested the feasibility of using intramyocardial injection or intracoronary infusions of recombinant adeno-a ... | 1999 | 9892583 |
| optimization of packaging of adeno-associated virus gene therapy vectors using plasmid transfections. | adeno-associated virus (aav) is attracting wide attention as a potential human gene therapy vector. the advantages of this vector system are that it is naturally defective, it readily integrates into the target cell's genome and is considered to be nonpathogenic. aav infects a wide variety of cell and tissue types. the major disadvantages of the vector are its small size and the labor-intensive procedures required to prepare large amounts of the vector for clinical studies. in this manuscript we ... | 1998 | 9923737 |
| efficient coexpression and secretion of anti-atherogenic human apolipoprotein ai and lecithin-cholesterol acyltransferase by cultured muscle cells using adeno-associated virus plasmid vectors. | plasma apolipoprotein ai (apoai) and lecithin-cholesterol acyltransferase (lcat) play important roles in reverse cholesterol transport, promoting the removal of excess cholesterol from peripheral cells and reducing formation of atherosclerotic lesions. gene augmentation of either apoai or lcat, or both, are thus attractive targets for prevention or treatment of atherosclerosis. with the eventual aim of safe and efficient gene delivery to skeletal muscle, our chosen secretory platform for systemi ... | 1998 | 9930350 |
| antisense inhibition of at1 receptor in vascular smooth muscle cells using adeno-associated virus-based vector. | vascular smooth muscle cells (vsmcs) are the main peripheral target for vasoconstriction and growth-promoting activity of angiotensin ii (ang ii), acting through angiotensin type 1 receptors (at1-r). current antihypertension treatments include daily reductions in the effects of ang ii. to decrease an effect of ang ii in a prolonged fashion, we have developed an adeno-associated virus (aav) vector with antisense dna for at1-r. aav has many advantages over other viral vectors. aav is nonpathogenic ... | 1999 | 9931129 |
| gene therapy for cerebrovascular disease. | to review the principles of and the experimental and clinical results of gene therapy for cerebrovascular disease. | 1999 | 9932877 |
| 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 |
| analysis of the effects of charge cluster mutations in adeno-associated virus rep68 protein in vitro. | the rep78 and rep68 proteins of adeno-associated virus type 2 (aav) are multifunctional proteins which are required for viral replication, regulation of aav promoters, and preferential integration of the aav genome into a region of human chromosome 19. these proteins bind the hairpin structures formed by the aav inverted terminal repeat (itr) origins of replication, make site- and strand-specific endonuclease cuts within the aav itrs, and display nucleoside triphosphate-dependent helicase activi ... | 1999 | 9971790 |
| 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 |
| neonatal intramuscular injection with recombinant adeno-associated virus results in prolonged beta-glucuronidase expression in situ and correction of liver pathology in mucopolysaccharidosis type vii mice. | for many metabolic diseases, early correction of the inherited deficiency is required to prevent long-term sequelae. we examined the ability of adeno-associated virus (aav) to mediate efficient gene transfer during the neonatal period in mice with the lysosomal storage disease mucopolysaccharidosis type vii (mps vii). quadriceps of newborn mps vii mice were injected with an aav vector containing human beta-glucuronidase (gusb) cdna. high-level intramuscular gusb expression was seen as early as 2 ... | 1999 | 10022533 |
| adeno-associated virus-mediated gene transfer to the brain: duration and modulation of expression. | adeno-associated virus (aav) is a promising vector for central nervous system (cns) gene transfer, but a number of issues must be addressed if aav is to be used for widespread delivery throughout the cns. our aim was to test the effect of dose, route of delivery, and hydroxyurea treatment on brain expression of beta-galactosidase activity after cerebral inoculation with an raav-lacz vector (raav-beta-gal). we also wished to test whether an immune response appeared against the vector and the tran ... | 1999 | 10022545 |
| recombinant adeno-associated virus-mediated expression of o6-alkylguanine-dna-alkyltransferase protects human epithelial and hematopoietic cells against chloroethylating agent toxicity. | recombinant adeno-associated virus (raav) encoding the human o6-alkylguanine-dna-alkyltransferase (hat) protein and a selectable marker (neo(r)) was used to transduce human cervical carcinoma (hela) cells and erythroleukemic (k562) cells and clones were selected using g418 (0.4 mg/ml). thirteen hela clones were isolated, 9 of which survived for 2-3 months before cell death ensued, presumably owing to the loss of g418 resistance. northern blot analysis of the remaining four clones, using a neo pr ... | 1999 | 10022554 |
| inducible long-term gene expression in brain with adeno-associated virus gene transfer. | recombinant adeno-associated virus (raav) vectors hold promise for treating a number of neurological disorders due to the ability to deliver long-term gene expression without toxicity or immune response. critical to these endeavors will be controlled expression of the therapeutic gene in target cells. we have constructed and tested a dual cassette raav vector carrying a reporter gene under the control of the tetracycline-responsive system and the tetracycline transactivator. transduction in vitr ... | 1998 | 10023439 |
| treatment of lysosomal storage disease in mps vii mice using a recombinant adeno-associated virus. | mucopolysaccharidosis type vii (mps vii) is a lysosomal storage disease caused by a genetic deficiency of beta-glucuronidase (gus). we used a recombinant adeno-associated virus vector (aav-gus) to deliver gus cdna to mps vii mice. the route of vector administration had a dramatic effect on the extent and distribution of gus activity. intramuscular injection of aav-gus resulted in high, localized production of gus, while intravenous administration produced low gus activity in several tissues. thi ... | 1998 | 10023443 |
| human factor viii can be packaged and functionally expressed in an adeno-associated virus background: applicability to haemophilia a gene therapy. | adeno-associated virus (aav) is a single-stranded dna parvovirus displaying several attractive features applicable to haemophilia a gene therapy, including nonpathogenicity and potential for long-term transgene expression from either integrated or episomal forms. we have generated and characterized two b-domain-deleted (bdd) fviii mutants, deleted in residues phe756 to ile1679 (fviiidelta756-1679) or thr761 to asn1639 (fviiidelta761-1639). [35s]metabolic labelling experiments and immunoprecipita ... | 1999 | 10027708 |
| 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 |
| neonatal gene transfer leads to widespread correction of pathology in a murine model of lysosomal storage disease. | for many inborn errors of metabolism, early treatment is critical to prevent long-term developmental sequelae. we have used a gene-therapy approach to demonstrate this concept in a murine model of mucopolysaccharidosis type vii (mps vii). newborn mps vii mice received a single intravenous injection with 5.4 x 10(6) infectious units of recombinant adeno-associated virus encoding the human beta-glucuronidase (gusb) cdna. therapeutic levels of gusb expression were achieved by 1 week of age in liver ... | 1999 | 10051635 |
| 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 |
| transduction of primitive human marrow and cord blood-derived hematopoietic progenitor cells with adeno-associated virus vectors. | we evaluated the capacity of adeno-associated virus (aav) vectors to transduce primitive human myeloid progenitor cells derived from marrow and cord blood in long-term cultures and long-term culture-initiating cell (ltc-ic) assays. single-colony analyses showed that aav vectors transduced cd34(+) and cd34(+)38(-) clonogenic cells in long-term culture. gene transfer was readily observed in ltc-ics derived from 5-, 8-, and 10-week cultures. recombinant aav (raav) transduction was observed in every ... | 1999 | 10068661 |
| charged-to-alanine scanning mutagenesis of the n-terminal half of adeno-associated virus type 2 rep78 protein. | the adeno-associated virus (aav) rep78 and rep68 proteins are required for site-specific integration of the aav genome into the aavs1 locus (19q13.3-qter) as well as for viral dna replication. rep78 and rep68 bind to the gagc motif on the inverted terminal repeat (itr) and cut at the trs (terminal resolution site). a similar reaction is believed to occur in aavs1 harboring an analogous gagc motif and a trs homolog, followed by integration of the aav genome. to elucidate the functional domains of ... | 1999 | 10074114 |
| comparative characterization of rep proteins from the helper-dependent adeno-associated virus type 2 and the autonomous goose parvovirus. | adeno-associated viruses (aavs) are nonautonomous human parvoviruses in that they are dependent on helper functions supplied by other viruses or on genotoxic stimuli for conditions permissive for replication. in the absence of helper, aav type 2 enters latency by integration into a specific site on human chromosome 19. this feature of aav, in combination with a lack of pathogenicity, makes aav an attractive candidate vector for human gene therapy. goose parvovirus (gpv) is both autonomous and pa ... | 1999 | 10074142 |
| disease-inducible transgene expression from a recombinant adeno-associated virus vector in a rat arthritis model. | rheumatoid arthritis (ra) is a systemic autoimmune disease affecting 1% of the world's population, with significant morbidity and mortality. in this study, we investigated a recombinant adeno-associated virus (raav) vector for its potential application in ra gene therapy. raav encoding escherichia coli beta-galactosidase was injected into rat joints which had already been induced into acute arthritis after local lipopolysaccharide (lps) administration, and the efficiency of in vivo transduction ... | 1999 | 10074195 |
| site-specific integration mediated by a hybrid adenovirus/adeno-associated virus vector. | adenovirus (ad) and adeno-associated virus (aav) have attractive and complementary properties that can be exploited for gene transfer purposes. ad vectors are probably the most efficient vehicles to deliver foreign genes both in vitro and in vivo. aav exhibits the unique ability to establish latency by efficiently integrating at a specific locus of human chromosome 19 (aavs1). two viral elements are necessary for the integration at aavs1: rep68/78 and the inverted terminal repeats (aav-itrs). in ... | 1999 | 10077559 |
| reversal of diabetes in streptozotocin-treated rats by intramuscular injection of recombinant adeno-associated virus containing rat preproinsulin ii gene. | 1999 | 10083275 | |
| reversal of diabetes in the rat by injection of hematopoietic stem cells infected with recombinant adeno-associated virus containing the preproinsulin ii gene. | 1999 | 10083298 | |
| nontargeted stable integration of recombinant adeno-associated virus into human leukemia and lymphoma cell lines as evaluated by fluorescence in situ hybridization. | a number of studies on human epithelial cells of varying origin have demonstrated integration of recombinant adeno-associated virus (aav) vectors into a variety of chromosomes compared with the site-specific integration on chromosome 19 predominantly observed for wild-type (wt) aav. we have constructed a recombinant aav (raav) vector and tested the integration into hematopoietic cells, using the human acute myeloid leukemia cell line aml5 and the human non-hodgkin's lymphoma cell line oci-ly18 a ... | 1999 | 10094197 |
| two independent molecular pathways for recombinant adeno-associated virus genome conversion occur after uv-c and e4orf6 augmentation of transduction. | numerous environmental influences have been demonstrated to enhance recombinant adeno-associated virus (raav) transduction. such findings are the foundation of developing new and innovative strategies to improve the efficiency of raav as a gene therapy vector. several of these environmental factors included genotoxic stresses such as uv and y irradiation as well as certain adenoviral gene products such as e4orf6. the mechanisms by which these environmental stimuli increase raav transduction are ... | 1999 | 10094202 |
| evaluation of adeno-associated virus-mediated gene transfer into the rat retina by clinical fluorescence photography. | the purpose of this study was to evaluate recombinant adeno-associated virus (aav) as an in vivo gene transfer vector for the retina and to explore the possibility of monitoring the expression of green fluorescent protein (gfp) using a noninvasive method. rats were injected subretinally with raav-gfp or raav-lacz. strong expression of the reporter gene in a circular area surrounding the injection site was observed in retinal whole mounts and tissue sections. higher magnification revealed that ce ... | 1999 | 10094207 |
| enhancement of uv-induced cytotoxicity by the adeno-associated virus replication proteins. | adeno-associated virus (aav) normally requires co-infection of a helper virus to complete its life cycle. however, under conditions of cellular stress, such as treatment with carcinogens or ultraviolet (uv) light, a permissive intracellular environment is established and aav completes its replicative cycle producing low levels of progeny virus. aav dna replication is dependent upon viral replication proteins, rep78 and rep68. the detailed mechanism by which these proteins interact with host cell ... | 1999 | 10095060 |
| sustained correction of bleeding disorder in hemophilia b mice by gene therapy. | mice generated by disrupting the clotting factor ix gene exhibit severe bleeding disorder and closely resemble the phenotype seen in hemophilia b patients. here we demonstrate that a single intraportal injection of a recombinant adeno-associated virus (aav) vector encoding canine factor ix cdna under the control of a liver-specific enhancer/promoter leads to a long-term and complete correction of the bleeding disorder. high level expression of up to 15-20 microgram/ml of canine factor ix was det ... | 1999 | 10097136 |
| involvement of protein-dna interaction in adeno-associated virus rep78-mediated inhibition of hiv-1. | it has been well documented by several laboratories that adeno-associated virus (aav) is able to inhibit hiv-1 replication and gene expression. this effect has been mapped to the aav-encoded rep78 protein. however, the mechanism by which rep78 is able to inhibit hiv-1 is unclear. as rep78 is a dna binding transcription factor, the objective of this study was to investigate where rep78 might bind within the hiv-1 long terminal repeat (ltr) sequences and to judge the importance of this protein-dna ... | 1998 | 10195265 |
| recombinant adeno-associated virus-based vectors provide short-term rather than long-term transduction of primitive hematopoietic stem cells. | bone marrow stem cells collected from b6-gpi-1a mice pretreated with 5-fluorouracil were incubated for 2 h at 37 degrees c in the presence of the recombinant adenovirus-associated virus-based vector (raav) ssv9. as measured in vitro immediately following transduction, ssv9 was found to be effective in transducing the primitive cobble-stone-area-forming cell (cafc)-35 subset (60% transduction efficiency). however, this did not predict long-term expression as the presence of the transgene could no ... | 1999 | 10195572 |
| gene therapy vectors based on adeno-associated virus type 1. | the complete sequence of adeno-associated virus type 1 (aav-1) was defined. its genome of 4,718 nucleotides demonstrates high homology with those of other aav serotypes, including aav-6, which appears to have arisen from homologous recombination between aav-1 and aav-2. analysis of sera from nonhuman and human primates for neutralizing antibodies (nab) against aav-1 and aav-2 revealed the following. (i) nab to aav-1 are more common than nab to aav-2 in nonhuman primates, while the reverse is tru ... | 1999 | 10196295 |
| adeno-associated virus (aav) type 5 rep protein cleaves a unique terminal resolution site compared with other aav serotypes. | adeno-associated virus (aav) replication depends on two viral components for replication: the aav nonstructural proteins (rep) in trans, and inverted terminal repeat (itr) sequences in cis. aav type 5 (aav5) is a distinct virus compared to the other cloned aav serotypes. whereas the rep proteins and itrs of other serotypes are interchangeable and can be used to produce recombinant viral particles of a different serotype, aav5 rep proteins cannot cross-complement in the packaging of a genome with ... | 1999 | 10196327 |
| functional expression of exogenous proteins in mammalian sensory hair cells infected with adenoviral vectors. | to understand the function of specific proteins in sensory hair cells, it is necessary to add or inactivate those proteins in a system where their physiological effects can be studied. unfortunately, the usefulness of heterologous expression systems for the study of many hair cell proteins is limited by the inherent difficulty of reconstituting the hair cell's exquisite cytoarchitecture. expression of exogenous proteins within hair cells themselves may provide an alternative approach. because re ... | 1999 | 10200223 |
| 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 |
| enhanced sensitivity of small cell lung cancer cell lines to cisplatin and etoposide after infection with adeno-associated virus type 2. | in previous studies we have reported the sensitisation of human tumour cells to gamma irradiation and chemotherapeutic drugs upon infection with the human non-pathogenic adeno-associated virus type 2 (aav-2) in vitro and in vivo. treatment of small cell lung cancer (sclc) is consistently hampered by relapses due to the selection of chemotherapy-resistant cell clones. hence, we were interested to test whether selection of chemotherapy-resistant sclc cells might be reduced or even prevented if che ... | 1999 | 10211097 |
| long-term actions of vector-derived nerve growth factor or brain-derived neurotrophic factor on choline acetyltransferase and trk receptor levels in the adult rat basal forebrain. | trophic factor gene therapy may provide a rational treatment strategy for neurodegenerative disease. recombinant adeno-associated virus vectors, incorporating a neuron-specific promoter driving bicistronic expression of green fluorescent protein and either nerve growth factor or brain-derived neurotrophic factor, transduced 10,000-15,000 neurons in the medial septum for periods of at least six months. both cholinergic and non-cholinergic neurons expressed green fluorescent protein. nerve growth ... | 1999 | 10218782 |
| highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses. | gene transfer vectors based on the replication-defective human parvovirus, adeno-associated virus type 2 (aav-2), are viable candidates for in vivo and ex vivo human use. however, widespread testing of aav vectors has been limited by difficulties in generating pure, high-titer vector stocks that are fully characterized. to address these issues, we have developed a single-step purification scheme using heparin affinity chromatography. recovery from the crude lysate starting material exceeds 70%, ... | 1999 | 10223736 |
| adeno-associated virus 2 co-receptors? | 1999 | 10229209 | |
| 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 |
| a 110-kda nuclear shuttle protein, nucleolin, specifically binds to adeno-associated virus type 2 (aav-2) capsid. | a 110-kda protein was copurified with adeno-associated virus type 2 (aav-2) virions after cscl density gradient isopycnic centrifugation. amino acid sequence of peptides derived from this protein after tryptic digestion, monoclonal antibody production, and western blot analysis showed that the copurified protein was the major nucleolar phosphoprotein, human nucleolin. virus overlay assays demonstrated that aav-2 capsid specifically bound to the human nucleolin, and immunoprecipitation studies co ... | 1999 | 10329548 |
| raav vector-mediated sarcogylcan gene transfer in a hamster model for limb girdle muscular dystrophy. | the limb girdle muscular dystrophies (lgmd) are a genetically and phenotypically heterogeneous group of degenerative neuromuscular diseases. a subset of the genetically recessive forms of lgmd are caused by mutations in the four muscle sarcoglycan genes (alpha, beta, gamma and delta). the coding sequences of all known sarcoglycan genes are smaller than 2 kb, and thus can be readily packaged in recombinant adeno-associated virus (raav) vectors. previously, we have demonstrated highly efficient an ... | 1999 | 10341878 |
| hepatic gene therapy using adeno-associated virus vectors. | recombinant adeno-associated virus vectors have been shown to safely transduce a number of tissues in preclinical animal studies. the level of gene transfer is sufficient to successfully treat a large number of medical disorders. moreover, the long-term persistence of the vector sequences in animals makes it likely that this vector will be useful for genetic diseases requiring life-long therapy. this review outlines the biological principles of the vector, as well as its advantages and current l ... | 1999 | 10349684 |
| ca2+ and phorbol ester effect on the mast cell phosphoprotein induced by cromolyn. | several phosphoproteins are involved in stimulus-secretion coupling. the beta and gamma subunits of immunoglobulin e binding protein (fc epsilonri) and three other protein bands get phosphorylated during stimulation of mast cell secretion. these additional proteins of 42, 59 and 68 kda are also phosphorylated when secretion is stimulated by compound 48/80 (c48/80). a 78 kda band, however, is phosphorylated as secretion wanes after stimulation with c48/80 and by the anti-allergic drug disodium cr ... | 1999 | 10357262 |
| efficient production of adeno-associated virus vectors using split-type helper plasmids. | adeno-associated virus (aav) vectors are potentially useful vehicles for the delivery of therapeutic genes into human cells. to determine the optimal expression pattern of aav proteins (rep78, rep68, rep52, rep40, and cap proteins) for packaging the recombinant aav genome, helper plasmids were split into two portions. in this study, two sets of split-type helper plasmids were prepared; i.e., 1) a rep expression plasmid (prep) and cap expression plasmid (pcap), and 2) a large rep expression plasm ... | 1999 | 10363588 |
| isolation of recombinant adeno-associated virus vector-cellular dna junctions from mouse liver. | recombinant adeno-associated virus (raav) vectors allow for sustained expression of transgene products from mouse liver following a single portal vein administration. here a raav vector expressing human coagulation factor f.ix (hf.ix), aav-ef1alpha-f.ix (hf.ix expression was controlled by the human elongation factor 1alpha [ef1alpha] enhancer-promoter) was injected into mice via the portal vein or tail vein, or directly into the liver parenchyma, and the forms of raav vector dna extracted from t ... | 1999 | 10364291 |
| transduction of well-differentiated airway epithelium by recombinant adeno-associated virus is limited by vector entry. | the limitations of adeno-associated virus (aav)-mediated vectors for lung-directed gene transfer were investigated by using differentiated human respiratory epithelium in air-liquid interface cultures. transduction efficiency was high in undifferentiated cells and was enhanced in well-differentiated cells after basolateral application of the vector or after apical application following disruption of tight junctions or pretreatment of the cultures with glycosidases. these results indicate that tr ... | 1999 | 10364362 |
| suppression of tumorigenicity in cervical carcinoma hela cells by an episomal form of adeno-associated virus. | to compare the oncosuppressive activity of integrated and episomal adeno-associated virus (aav), two epstein-barr virus (ebv) derived episomal form vectors, p205 and p220, were used to generate plasmids containing episomal aav. hela cells transfected with p205, p220, and the plasmids with forward (pa205-1, pa220-1) and reverse (pa205-1, pa220-2) orientation of inserted aav dna were designated h205, h220, ha205-1, ha220-1, ha205-1, and ha220-2, respectively. the respective average copy numbers of ... | 1999 | 10375600 |
| generation of aberrant sprouting in the adult rat brain by gap-43 somatic gene transfer. | the expression of gap-43 was modulated genetically in the adult rat nigrostriatal or septohippocampal pathway using recombinant adeno-associated virus (raav) vectors incorporating the neuron specific enolase (nse) promoter and either a rat gap-43 cdna or the corresponding antisense sequence. bicistronic expression of green fluorescent protein (gfp) enabled us to evaluate transduced neurons selectively. single injections of raav into the substantia nigra pars compacta (snc) transduced both dopami ... | 1999 | 10375659 |
| exogenous control of cardiac gene therapy: evidence of regulated myocardial transgene expression after adenovirus and adeno-associated virus transfer of expression cassettes containing corticosteroid response element promoters. | because of the relative inaccessibility of the heart for repeated gene therapy, it would be useful to regulate the expression of transgenes delivered in a single dose of a gene therapy vector. incorporation into the vector of a regulatable promoter that is responsive to pharmacologic agents that are widely used and well tolerated in clinical practice represents such a control strategy. | 1999 | 10384181 |
| [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 |
| [detection of ttv dna in hepatocellular carcinoma]. | a new hepatitis agent, ttv has been cloned from post-transfusion hepatitis patient sera. this virus has a single stranded dna as genome, surrounded by viral capsid antigen without envelope. it might be a member of parvoviridae. but genome organization is far from the known parvoviridae such as b19 and adeno-associated virus. although this virus could be a new causative agent for nona-nong hepatitis, high healthy carrier rate, no relation with hepatocellular carcinoma and no evidence of amplifica ... | 1999 | 10391002 |
| long-term restoration of striatal l-aromatic amino acid decarboxylase activity using recombinant adeno-associated viral vector gene transfer in a rodent model of parkinson's disease. | as a potential treatment for parkinson's disease, viral vector-mediated over-expression of striatal l-aromatic amino acid decarboxylase was tested in an attempt to facilitate the production of therapeutic levels of dopamine after peripheral l-dihydroxyphenylalanine administration. the results of microdialysis and enzyme activity assays indicate that striatal decarboxylation of peripherally administered l-dihydroxyphenylalanine was enhanced by recombinant adeno-associated virus-mediated gene tran ... | 1999 | 10392841 |
| persistent transgene product in retina, optic nerve and brain after intraocular injection of raav. | recombinant adeno-associated virus (raav) is a promising vector for retinal application as it transduces photoreceptors and retinal pigment epithelium cells efficiently and in a stable fashion. because raav also transduces retinal ganglion cells, we reasoned that ocular application of raav might result in delivery of transgenic protein to the cns. here we describe high levels of green fluorescent protein (gfp) persisting at least 6 months in optic nerves and brains of mice and dogs after intravi ... | 1999 | 10396623 |
| recombinant adeno-associated virus (aav) drives constitutive production of glutamate decarboxylase in neural cell lines. | many neurological disorders result directly or indirectly from the loss of inhibitory function. engineering the production of gaba, an inhibitory neurotransmitter, may therefore be able at least partly to restore the lost inhibition seen in epilepsy, parkinson's disease, or huntington's disease. in this article, we describe a set of recombinant adeno-associated viruses (aavs) that can deliver cdnas encoding the gaba-producing enzyme, glutamate decarboxylase (gad), directly into neural cells. we ... | 1999 | 10397644 |
| reporter expression persists 1 year after adeno-associated virus-mediated gene transfer to the optic nerve. | to determine the foci and duration of protein expression following virus-mediated gene transfer to the optic nerve. | 1999 | 10408459 |
| long-term regulated expression of growth hormone in mice after intramuscular gene transfer. | effective delivery of secreted proteins by gene therapy will require a vector that directs stable delivery of a transgene and a regulatory system that permits pharmacologic control over the level and kinetics of therapeutic protein expression. we previously described a regulatory system that enables transcription of a target gene to be controlled by rapamycin, an orally bioavailable drug. here we demonstrate in vivo regulation of gene expression after intramuscular injection of two separate aden ... | 1999 | 10411931 |
| induction of immunity to antigens expressed by recombinant adeno-associated virus depends on the route of administration. | recombinant adeno-associated virus (raav) is a replication-defective parvovirus which is being explored as a vector for gene therapy because of its broad host range, excellent safety profile, and durable transgene expression in infected hosts. raav has also been reported by several groups to induce little or no immune response to its encoded transgene products. in this study we examined the immunogenicity of raav by studying the immune response of c57bl/6 mice to a single dose of raav-encoding o ... | 1999 | 10413654 |
| adeno-associated virus vectors and hematology. | 1999 | 10419876 | |
| 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 |
| 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 |
| transfer of activation-dependent gene expression into t cell lines by recombinant adeno-associated virus. | we examined the ability of recombinant adeno-associated virus (raav) to transfer regulated gene expression into t cell lines. an aav-based vector containing the neomycin resistance gene and expressing the firefly luciferase (luc) gene under the regulatory control of the interleukin 2 promoter (paav-luc) was generated and adenovirus-free raav (raav-luc) was produced from this vector. transfection of paav-luc into the human t cell line jurkat resulted in luciferase expression while infection of ju ... | 1999 | 10435102 |
| production of recombinant adeno-associated virus vectors using a packaging cell line and a hybrid recombinant adenovirus. | recombinant adeno-associated virus (raav) vectors are under consideration for a wide variety of gene therapy applications. one of the limitations of the raav vector system has been the difficulty in producing the vector in sufficient quantity for adequate preclinical and clinical evaluation. a common method for vector production involves large-scale transient transfection of multiple plasmids into cultured cells. because this approach might not be feasible for clinical scale manufacturing, we ha ... | 1999 | 10435114 |
| 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 |
| nuclear transport of the major capsid protein is essential for adeno-associated virus capsid formation. | adeno-associated virus capsids are composed of three proteins, vp1, vp2, and vp3. although vp1 is necessary for viral infection, it is not essential for capsid formation. the other capsid proteins, vp2 and vp3, are sufficient for capsid formation, but the functional roles of each protein are still not well understood. by analyzing a series of deletion mutants of vp2, we identified a region necessary for nuclear transfer of vp2 and found that the efficiency of nuclear localization of the capsid p ... | 1999 | 10438891 |
| adeno-associated virus and development of cervical neoplasia. | evidence from several sources has suggested that adeno-associated virus (aav) infection might protect against cervical cancer, in part, by interfering with human papillomavirus (hpv)-induced tumorigenesis. detection of aav type 2 (aav-2) dna in cervical tissues has been reported. however, there have been few in vivo studies of women with cervical hpv infection or neoplasia, and these have reported inconsistent results. therefore, we used polymerase chain reaction (pcr) assays targeted to the aav ... | 1999 | 10440809 |
| application of gene therapy to acute inflammatory diseases. | the application of gene therapy to acute inflammation has not received as much research attention as has the treatment of genetically-based diseases, cancer, and viral infections. however, gene therapy as a drug delivery system offers several theoretical and practical advantages over current protein delivery systems. these include the ability to target therapies to individual tissues or cell types, to locally produce proteins that can act intracellularly or in an autocrine, juxtacrine, or paracr ... | 1999 | 10446888 |
| purification of recombinant adeno-associated virus by iodixanol gradient ultracentrifugation allows rapid and reproducible preparation of vector stocks for gene transfer in the nervous system. | recombinant adeno-associated virus (raav) vectors have become attractive tools for in vivo gene transfer. the production and purification of high-titer raav vector stocks for experimental and therapeutic gene transfer continue to undergo improvement. standard raav vector purification protocols include the purification of the vector by cesium chloride (cscl)-density gradient centrifugation followed by extensive desalination via dialysis against a physiological buffer for in vivo use. these proced ... | 1999 | 10446928 |
| interaction of adeno-associated virus rep78 with p53: implications in growth inhibition. | adeno-associated virus (aav) is a nonpathogenic, single-stranded dna virus belonging to the parvoviridae family. onco-suppressive properties of aav against adenovirus, a dna tumor virus, have been well documented. rep78, a major regulatory protein of aav, is believed to be responsible for its antioncogenic properties. most dna tumor viruses disturb the cell cycle pathways by essentially abrogating the functions of p53. here we present evidence that aav acts as an antiproliferative agent against ... | 1999 | 10446967 |
| stable transgene expression in rod photoreceptors after recombinant adeno-associated virus-mediated gene transfer to monkey retina. | recombinant adeno-associated virus (raav) is a promising vector for therapy of retinal degenerative diseases. we evaluated the efficiency, cellular specificity, and safety of retinal cell transduction in nonhuman primates after subretinal delivery of an raav carrying a cdna encoding green fluorescent protein (egfp), raav. cmv.egfp. the treatment results in efficient and stable egfp expression lasting >1 year. transgene expression in the neural retina is limited exclusively to rod photoreceptors. ... | 1999 | 10449795 |
| 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 |
| adeno-associated virus as a gene delivery vector for liver cells. | 1999 | 10453960 | |
| recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. | conventional methods for raav purification that are based on cesium chloride ultracentrifugation have often produced vector preparations of variable quality and resulted in significant loss of particle infectivity. we report here several novel purification strategies that involve the use of non-ionic iodixanol gradients followed by ion exchange or heparin affinity chromatography by either conventional or hplc columns. these methods result in more than 50% recovery of raav from a crude lysate and ... | 1999 | 10455399 |
| high-titer recombinant adeno-associated virus production utilizing a recombinant herpes simplex virus type i vector expressing aav-2 rep and cap. | recombinant adeno-associated virus type 2 (raav) vectors have recently been used to achieve long-term, high level transduction in vivo. further development of raav vectors for clinical use requires significant technological improvements in large-scale vector production. in order to facilitate the production of raav vectors, a recombinant herpes simplex virus type i vector (rhsv-1) which does not produce icp27, has been engineered to express the aav-2 rep and cap genes. the optimal dose of this v ... | 1999 | 10455400 |
| cellular contaminants of adeno-associated virus vector stocks can enhance transduction. | transduction efficiency of different types of recombinant (r)aav-2 based vectors preparations markedly differed, with apparently no correlation with the replicative titers. using hela cells as target for transduction, 105 and 30 infectious units were necessary to observe one transductant using respectively cesium-chloride-purified raav and crude lysates of producer cells obtained by sonication. the purified vectors were however able to transduce hek-193 cells efficiently, but transgene expressio ... | 1999 | 10455407 |
| elimination of lysosomal storage in brains of mps vii mice treated by intrathecal administration of an adeno-associated virus vector. | mucopolysaccharidosis type vii (mps vii) is an inherited lysosomal storage disease caused by insufficient beta-glucuronidase (gus). to provide gene therapy in a mutant mouse model of this disease, we have used a recombinant adeno-associated virus (raav) vector to deliver gus cdna to a variety of tissues. although intravenous administration of vector produced therapeutic levels of gus in the liver, delivery to the brain was inadequate. to improve delivery to the brain intrathecal injection of the ... | 1999 | 10455422 |
| recombinant aav-2 harboring gfp-antisense/ribozyme fusion sequences monitor transduction, gene expression, and show anti-hiv-1 efficacy. | vector-mediated delivery of potentially antivirally active genes is a key step in somatic gene therapy including therapeutic approaches against aids. a crucial technical prerequisite is to monitor dna transfer into target cells. here, we describe recombinant infectious particles derived from the adeno-associated virus type 2 (aav-2) that are suitable to deliver effective hiv-1-directed antisense and ribozyme genes into target cells. to monitor transduction, we designed and tested a number of fus ... | 1999 | 10455431 |