Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
|---|
| control of adeno-associated virus type 2 cap gene expression: relative influence of helper virus, terminal repeats, and rep proteins. | adeno-associated virus type 2 (aav-2) gene expression is tightly controlled by functions of the helper virus as well as by the products of its own viral rep gene. double-immunofluorescence studies of rep and vp protein expression in cells coinfected with aav-2 and adenovirus type 2 showed that a large proportion of these cells expressed rep78 and rep52 but no capsid proteins. the percentage of rep78/rep52- and capsid protein-positive cells was strongly influenced by the relative ratio of aav-2 t ... | 1997 | 9343200 |
| 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 |
| 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 |
| a novel 165-base-pair terminal repeat sequence is the sole cis requirement for the adeno-associated virus life cycle. | adeno-associated virus (aav) replication is dependent on two copies of a 145-bp inverted terminal repeat (itr) that flank the aav genome. this is the primary cis-acting element required for productive infection and the generation of recombinant aav (raav) vectors. we have engineered a plasmid (pdd-2) containing only 165 bp of aav sequence: two copies of the d element, a unique sequence adjacent to the aav nicking site, flanking a single itr. when assayed in vivo, this modified hairpin was suffic ... | 1997 | 8995611 |
| the adeno-associated virus (aav) rep protein acts as both a repressor and an activator to regulate aav transcription during a productive infection. | adeno-associated virus (aav) uses three promoters, p5, p19, and p40, to regulate viral gene expression. the p5 and p19 promoters direct the synthesis of the viral regulatory proteins, rep78 and -68 and rep52 and -40, respectively. the p5 rep proteins bind a linear 22-bp sequence, the rep binding element (rbe), that is within both the terminal repeat (tr) and the p5 promoter. in the absence of helper virus, all four rep proteins have been shown to reduce transcription from the viral p5 and p19 pr ... | 1997 | 8995628 |
| a novel terminal resolution-like site in the adeno-associated virus type 2 genome. | the adeno-associated virus 2 (aav) contains a single-stranded dna genome of which the terminal 145 nucleotides are palindromic and form t-shaped hairpin structures. these inverted terminal repeats (itrs) play an important role in aav dna replication and resolution, since each of the itrs contains a terminal resolution site (trs) that is the target site for the aav rep gene products (rep). however, the rep proteins also interact with the aav dna sequences that lie outside the itrs, and the itrs a ... | 1997 | 8995635 |
| subcellular compartmentalization of adeno-associated virus type 2 assembly. | using immunofluorescence and in situ hybridization techniques, we studied the intracellular localization of adeno-associated virus type 2 (aav-2) rep proteins, vp proteins, and dna during the course of an aav-2/adenovirus type 2 coinfection. in an early stage, the rep proteins showed a punctate distribution pattern over the nuclei of infected cells, reminiscent of replication foci. at this stage, no capsid proteins were detectable. at later stages, the rep proteins were distributed more homogene ... | 1997 | 8995658 |
| gene therapy for neurologic disease: benchtop discoveries to bedside applications. 1. the bench. | the overall goal of this review is to provide the pediatric neurologist with a theoretical foundation in gene therapy. gene therapy became feasible in the early 1970s and the first transfer of a foreign gene into humans was approved by the nih in 1989. adenovirus, adeno-associated virus, herpes-simplex virus, retroviruses, and other vectors have been used to efficiently transduce genes into cells in vitro and in vivo. we discuss laboratory experiments that have provided a strong scientific ratio ... | 1997 | 9010789 |
| the adeno-associated virus rep78 major regulatory protein forms multimeric complexes and the domain for this activity is contained within the carboxy-half of the molecule. | the adeno-associated virus (aav) encoded rep78 is a multifunctional protein which is able to regulate transcription, is required for aav dna replication, and appears necessary for site specific integration of aav dna into human chromosome 19. being analogous to the large t antigen, the replication protein of polyomaviruses which is known to homo-multimerize, it seemed likely that the rep78 protein would also interact with itself to carry out at least some of its functions. furthermore, in electr ... | 1997 | 9013883 |
| improved production of recombinant aav by transient transfection of nb324k cells using electroporation. | adeno-associated virus (aav) is useful as an integrating vector for gene transfer. aav recombinants are generally produced by transient co-transfection methods since it has proven difficult to generate stable packaging cell lines. acceptable titers of transducing recombinants should be obtainable by optimizing conditions for transient co-transfection. here, using a luciferase reporter derivative of the aav infectious plasmid psub201, we show that substantially higher yields of transducing virus ... | 1997 | 9015283 |
| comparison of retroviral and adeno-associated viral vectors designed to express human clotting factor ix. | several different designs for retroviral and adeno-associated virus (aav) vectors were developed to express human clotting factor ix. seven separate retroviral vectors were constructed, including chimeric long terminal repeat (ltr)-based designs, vectors containing splice donor/acceptor sites with internal ribosome entry sites (ires), and vectors with an internal cytomegalovirus (cmv)- or hepatitis b virus (hbv)-derived promoter. five aav vectors were produced using the same cassette design wher ... | 1997 | 9017417 |
| gene therapy for haematopoietic and lymphoid disorders. | gene transfer into haematopoietic stem cells (hsc) has been investigated for treatment of genetic disorders, conferral of chemotherapy resistance and insertion of genes to inhibit hiv-1 replication. methods have been available for almost a decade to transduce murine hsc using high-titre retroviral vectors and stimulation of hsc proliferation with cytokines such as il-3 and il-6. unfortunately, attempts to replicate the high efficiency of gene transfer using canine or simian gene transfer/bone ma ... | 1997 | 9020937 |
| the cellular transcription factor sp1 and an unknown cellular protein are required to mediate rep protein activation of the adeno-associated virus p19 promoter. | control of adeno-associated virus (aav) transcription from the three aav promoters (p5, p19, and p40) requires the adenovirus e1a protein and the aav nonstructural (rep) proteins. the rep proteins have been shown to repress the aav p5 promoter yet facilitate activation of the p19 and p40 promoters during a productive infection. to elucidate the mechanism of promoter regulation by the aav rep proteins, the cellular factors involved in mediating rep activation of the p19 promoter were characterize ... | 1997 | 9032303 |
| recombinant adeno-associated virus mediates a high level of gene transfer but less efficient integration in the k562 human hematopoietic cell line. | we tested the ability of a recombinant adeno-associated virus (raav) vector to express and integrate exogenous dna into human hematopoietic cells in the absence of selection. we developed an raav vector, aav-tngfr, carrying a truncated rat nerve growth factor receptor (tngfr) cdna as a cell surface reporter under the control of the moloney murine leukemia virus (momulv) long terminal repeat. an analogous momulv-based retroviral vector (l-tngfr) was used in parallel, and gene transfer and express ... | 1997 | 9032306 |
| analysis of recombinant adeno-associated virus packaging and requirements for rep and cap gene products. | adeno-associated virus (aav) is a human parvovirus currently being developed as a vector for gene therapy applications. because the gene transfer vector commonly retains only the aav terminal repeats, propagation of recombinant aav (raav) requires that the viral replication (rep) and capsid (cap) proteins be supplied in trans. in an effort to optimize the production of these vectors, a panel of helper plasmids was constructed to determine if expression of the rep and/or cap genes is a limiting f ... | 1997 | 9032320 |
| binding sites for adeno-associated virus rep proteins within the human genome. | the rep proteins of adeno-associated virus type 2 (aav) are known to bind to rep recognition sequences (rrss) in the aav inverted terminal repeats (itrs), the aav p5 promoter, and the preferred aav integration site in human chromosome 19, called aavs1. integration of the aav genome into aavs1 appears to be mediated by an interaction between the rep proteins of aav and rep binding sites within the viral genome and the integration locus. in an attempt to identify potential alternate integration si ... | 1997 | 9032395 |
| persistent expression of human clotting factor ix from mouse liver after intravenous injection of adeno-associated virus vectors. | we previously found that gene transduction by adeno-associated virus (aav) vectors in cell culture can be stimulated over 100-fold by treatment of the target cells with agents that affect dna metabolism, such as irradiation or topoisomerase inhibitors. here we show that previous gamma-irradiation increased the transduction rate in mouse liver by up to 900-fold, and the topoisomerase inhibitor etoposide increased transduction by about 20-fold. similar rates of hepatic transduction were obtained b ... | 1997 | 9037069 |
| antisense inhibition and adeno-associated viral vector delivery for reducing hypertension. | antisense oligodeoxynucleotides have been designed to inhibit the production of specific proteins. in models of hypertension, we have targeted the renin-angiotensin system at the level of synthesis (angiotensinogen) and the receptor (at1 receptor). the design of antisense oligonucleotides requires choosing a site to inhibit mrna processig or translation. the strategy we use is to make three oligonucleotides of antisense sequences, upstream and downstream from the aug site and over the aug site. ... | 1997 | 9039099 |
| toward cystic fibrosis gene therapy. | cystic fibrosis (cf) is a common genetic disorder characterized by defective epithelial chloride transport and progressive lung disease. although great strides have been made in the treatment of cf, it remains lethal, often by early adulthood. cf is one of the most extensively researched genetic diseases as a target for gene therapy development. it may also serve as an important model for gene therapy of other diseases. preclinical and clinical research has lead to the rapid development of a var ... | 1997 | 9046956 |
| lack of site-specific integration of the recombinant adeno-associated virus 2 genomes in human cells. | the adeno-associated virus 2 (aav)-based vector system has been suggested for its potential use in human gene therapy because the wild-type (wt) aav genome appears to integrate into the human chromosomal dna in a site-specific manner. we systematically investigated the integration patterns of the recombinant aav genomes lacking one or both the viral coding sequences. four recombinant aav genomes were constructed containing the genes for resistance to tetracycline (tcr) and the herpesvirus thymid ... | 1997 | 9048194 |
| hsv/aav hybrid amplicon vectors extend transgene expression in human glioma cells. | novel hybrid vectors, which incorporate critical elements of both herpes simplex virus type 1 (hsv-1) amplicon vectors and adeno-associated virus (aav) vectors, are able to sustain transgene expression in dividing glioma cells for over 2 weeks. these vectors combine the high infectibility and large transgene capacity of hsv-1 vectors with the potential for episomal amplification and chromosomal integration of aav vectors. the hybrid vectors contain the hsv-1 origin of dna replication, oris, and ... | 1997 | 9048203 |
| in vivo gene transfer into rat arterial walls with novel adeno-associated virus vectors. | we studied the ability of recombinant adeno-associated virus (raav) vectors to achieve gene transfer in vivo to intact rat carotid arteries. | 1997 | 9052570 |
| recombinant adeno-associated virus for muscle directed gene therapy. | although gene transfer with adeno-associated virus (aav) vectors has typically been low, transduction can be enhanced in the presence of adenovirus gene products through the formation of double stranded, non-integrated aav genomes. we describe the unexpected finding of high level and stable transgene expression in mice following intramuscular injection of purified recombinant aav (raav). the raav genome is efficiently incorporated into nuclei of differentiated muscle fibers where it persists as ... | 1997 | 9055858 |
| high-level globin gene expression mediated by a recombinant adeno-associated virus genome that contains the 3' gamma globin gene regulatory element and integrates as tandem copies in erythroid cells. | recombinant adeno-associated virus (raav) vectors are being evaluated for gene therapy applications. using purified raav containing a mutationally marked globin gene (a(gamma)*) and sites 2, 3, and 4 from the locus control region (rhs432a(gamma)*), but lacking a drug-resistance gene, we investigated the relationship between multiplicity of infection (moi), gene expression, and unselected genome integration in erythroid cells. most primary erythroid progenitors were transduced as reflected by a(g ... | 1997 | 9058741 |
| mutational analysis of the adeno-associated virus rep68 protein: identification of critical residues necessary for site-specific endonuclease activity. | the rep68 and rep78 proteins of adeno-associated virus type 2 (aav) are multifunctional proteins which contain overlapping amino acid sequences. they are required for viral replication and preferential integration of the aav genome into a region of human chromosome 19. during the terminal resolution process of aav dna replication, these proteins make a site-specific and strand-specific endonuclease cut within the aav inverted terminal repeat dna. the rep68 and rep78 proteins also have helicase a ... | 1997 | 9060625 |
| adeno-associated virus type 2 dna replication in vivo: mutation analyses of the d sequence in viral inverted terminal repeats. | the adeno-associated virus type 2 (aav) genome contains inverted terminal repeats (itrs) of 145 nucleotides. the terminal 125 nucleotides of each itr form palindromic hairpin (hp) structures that serve as primers for aav dna replication. these hp structures also play an important role in integration as well as rescue of the proviral genome from latently infected cells or from recombinant aav plasmids. each itr also contains a stretch of 20 nucleotides, designated the d sequence, that is not invo ... | 1997 | 9060669 |
| adeno-associated virus type 2-mediated transduction of murine hematopoietic cells with long-term repopulating ability and sustained expression of a human globin gene in vivo. | adeno-associated virus type 2 (aav), a nonpathogenic human parvovirus, is gaining attention as a vector for potential use in human gene therapy. we and others have described aav-mediated beta-globin gene transfer and expression in established human and murine erythroleukemia cell lines in vitro. however, successful aav-mediated globin gene transduction of hematopoietic stem cells and long-term expression in vivo in progeny cells have not been documented. we report here that infection of murine h ... | 1997 | 9060672 |
| adeno-associated virus rep78 protein and terminal repeats enhance integration of dna sequences into the cellular genome. | two adeno-associated virus (aav) elements are necessary for the integration of the aav genome: rep78/68 proteins and inverted terminal repeats (itrs). to study the contribution of the rep proteins and the itrs in the process of integration, we have compared the integration efficiencies of three different plasmids containing a green fluorescent protein (gfp) expression cassette. in one plasmid, no viral sequences were present; a second plasmid contained aav itrs flanking the reporter gene (integr ... | 1997 | 9060699 |
| site-specific integration by adeno-associated virus: a basis for a potential gene therapy vector. | 1997 | 9068788 | |
| efficient transduction of green fluorescent protein in spinal cord neurons using adeno-associated virus vectors containing cell type-specific promoters. | in this study, we have evaluated the capacity of recombinant adeno-associated virus (raav) vectors, containing cell type-specific promoters, to transduce neurons in vivo in the normal adult rat spinal cord. the neuron-specific enolase (nse) promoter and the platelet-derived growth factor (pdgf) b-chain promoter were used to direct expression of a 'humanized' form of the gene for green fluorescent protein (gfp). neuron-specific raavs were injected into the mid-cervical regions of adult rat spinal ... | 1997 | 9068791 |
| detection of adeno-associated virus type 2 sequences in the human genital tract. | adeno-associated virus (aav) is a defective parvovirus with unknown pathogenicity. it requires helper functions for its normal replication in human tissue and therefore is not readily isolated from clinical specimens. we have used the pcr method to examine the following clinical samples for the presence of aav sequences: (i) 15 nasopharyngeal aspirates from symptomatic patients, (ii) 7 swab or fluid specimens from vesicles of patients suspected of having varicella-zoster virus infections, (iii) ... | 1997 | 8968883 |
| adeno-associated virus as a gene delivery system. | adeno-associated virus (aav) has several characteristics which make it extremely attractive as a gene transfer vector: (1) no known pathogenicity; (2) high efficiency and the ability to remain latent; (3) a minimal number of antigens ensuring minimal immunogenicity; (4) the ability to transduce post-mitotic cells; (5) possible advantages of site-specific integration; and (6) a broad host and cell range. the human isolate, aav-2, is the best studied and has been the focus for gene delivery experi ... | 1997 | 10837553 |
| gene therapy for haemophilia. | progress toward the development of a gene therapy protocol for the treatment of haemophilia has been substantial. recent achievements include high level clotting factor expression in mice, dogs, and monkeys as well as phenotypic correction in both mouse and canine models of haemophilia. studies using adenoviral vectors have contributed to much of the recent success. however, the repertoire of gene transfer vehicles being applied to the development of gene therapy strategies for haemophilia has e ... | 1997 | 15989572 |
| new prospects for cardiovascular gene therapy. | cardiovascular gene therapy has been hampered by the lack of suitable gene delivery vectors for in vivo applications. low transduction efficiencies, lack of persistent transgene expression and undesirable inflammatory and immune responses have limited the prospects for human gene therapy in the cardiovascular system. new prospects for cardiovascular gene therapy are a result of recent vector developments, in particular with the use of adeno-associated virus (aav) based vectors in the heart and p ... | 1997 | 15989573 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| [the construction of a general adeno-associated virus vector]. | the general adeno-associated virus(aav) vector pacr-neo was constructed by substituting aav's construct gene with cassette that was composed of cmv-ie promoter, multiple cloning sites and sv40-polya. after transfecting pacr-neo into recombinant aav's packaging cell line ae1201, which was infected by adenovirus 5 before transfection, we got raav/acr-neo at the titer of 4.2 x 10(5) cfu/ml. using raav/acr-neo infected host cell a549, the recombinant aav could transfer reporter gene neo into host ce ... | 1998 | 12515194 |
| [adeno-associated virus vector mediated gene transfer of hsvi-tk and its effect on killing cancer cell]. | the plasmid pactk-19 was constructed by inserting hsvi-tk gene into the multiple cloning sites of the general adeno-associated virus(aav) vector pacr-neo. when plasmid pactk-19 was transfected to recombinant aav's packaging cell line ae1201, which was exposed to adenovirus 5 for two hours before transfection, we got raav/actk at the titer of 3.4 x 10(5) cfu/ml. after infecting human lung cancer cell a549 with raav/actk, we extracted host cell's chromosome cdna and amplified part of the hsvi-tk s ... | 1998 | 12526317 |
| cotransduction of tyrosine hydroxylase and aromatic l-amino acid decarboxylase genes into cultured striatal cells using adeno-associated virus vectors. | to examine whether tyrosine hydroxylase (th) and aromatic l-amino acid decarboxylase (aadc) genes can be cotransduced into the same target striatal cells using adeno-associated virus (aav) vectors, and to determine whether the cotransduction would result in better biochemical change than the th gene alone. | 1998 | 11263376 |
| infectious clones and vectors derived from adeno-associated virus (aav) serotypes other than aav type 2. | adeno-associated viruses (aavs) are single-stranded dependent parvoviruses being developed as transducing vectors. although at least five serotypes exist (aav types 1 to 5 [aav1 to -5]), only aav2, aav3, and aav4 have been sequenced, and the vectors in use were almost all derived from aav2. here we report the cloning and sequencing of a second aav3 genome and a new aav serotype designated aav6 that is related to aav1. aav2, aav3, and aav6 were 82% identical at the nucleotide sequence level, and ... | 1998 | 9420229 |
| role of the adenovirus dna-binding protein in in vitro adeno-associated virus dna replication. | a basic question in adeno-associated virus (aav) biology has been whether adenovirus (ad) infection provided any function which directly promoted replication of aav dna. previously in vitro assays for aav dna replication, using linear duplex aav dna as the template, uninfected or ad-infected hela cell extracts, and exogenous aav rep protein, demonstrated that ad infection provides a direct helper effect for aav dna replication. it was shown that the nature of this helper effect was to increase t ... | 1998 | 9420241 |
| adeno-associated virus type 2 vector for transduction of hematopoietic cells. | 1998 | 9304711 | |
| characterization of intrastriatal recombinant adeno-associated virus-mediated gene transfer of human tyrosine hydroxylase and human gtp-cyclohydrolase i in a rat model of parkinson's disease. | to achieve local, continuous l-dopa delivery in the striatum by gene replacement as a model for a gene therapy for parkinson's disease, the present studies used high titer purified recombinant adeno-associated virus (raav) containing cdnas encoding human tyrosine hydroxylase (hth) or human gtp-cyclohydrolase i [gtpchi, the rate-limiting enzyme for tetrahydrobiopterin (bh4) synthesis] or both to infect the 6-ohda denervated rat striatum. striatal th and gtpchi staining was observed 3 weeks after ... | 1998 | 9592104 |
| a high-throughput hybridization method for titer determination of viruses and gene therapy vectors. | one of the challenges facing researchers working with viruses and gene therapy vectors is the need to rapidly assay for infectious virus. current methods used to titer many viruses are cumbersome and are not amenable to handling large numbers of samples. here we describe the development of an assay that can rapidly quantify infectious viruses and gene therapy vectors. the assay relies on biological amplification of viral sequences and hybridization of labeled probes to immobilized nucleic acid f ... | 1998 | 9592173 |
| recombinant adeno-associated virus-mediated gene transfer into human leukemia cell lines. | adeno-associated virus (aav)-based vector is a promising gene transfer vehicle by virtue of the characteristics of wild-type aav:tropism to a wide range of human tissues and locus-specific integration at chromosome 19q13.3. to elucidate the nature of the recombinant aav (raav), transduction of neomycin phosphotransferase enzyme gene (neor gene) into seven human leukemia cell lines was performed. transduction efficiencies were assessed by colony formation assay and limiting dilution assay. the re ... | 1998 | 9594442 |
| expression of adeno-associated virus integrated transgene within the mammalian vestibular organs. | adeno-associated virus (aav) is a suitable viral vector for transgene expression within the mammalian vestibular organs. | 1998 | 9596192 |
| recombinant adeno-associated virus for the generation of autologous, gene-modified tumor vaccines: evidence for a high transduction efficiency into primary epithelial cancer cells. | to explore the potential of recombinant vectors based on recombinant adeno-associated virus (raav) for cancer vaccination, we investigated the transduction efficiency of raav into cancer cells ex vivo. infection of human epithelial cancer cell lines with raav carrying reporter genes encoding beta-galactosidase (raav/lacz) or luciferase (raav/luc) resulted in high levels of reporter gene expression (>90% positive cells). in marked contrast, raav poorly transduced all murine tumor cell lines, as w ... | 1998 | 9607416 |
| the adeno-associated virus rep78 major regulatory protein binds the cellular tata-binding protein in vitro and in vivo. | rep78 is the major regulatory protein of adenoassociated virus (aav). rep78 is able to transcriptionally regulate all three of aav's promoters, as well as a variety of heterologous promoters. in an attempt to understand the mechanism of action by which rep78 is able to regulate gene expression, we are investigating rep78's possible protein-protein interaction with basal transcription factors. one such critical basal transcription factor is the human tata binding protein, tbp. tbp is a core facto ... | 1998 | 9614873 |
| 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 |
| improvement of transduction efficiency of recombinant adeno-associated virus vector by entrapment in multilamellar liposomes. | recombinant adeno-associated virus (aav) has attracted considerable interest as a potential vector for human gene therapy, but its transduction efficiency is quite low. the present study demonstrated aav vector-associated liposomes to be more effective for in vitro gene transfer to human glioma cells than are liposomes containing plasmid dna. using vector-associated liposomes increased the transduction efficiency more than 10-fold compared to liposomes containing plasmid dna and more than 6-fold ... | 1998 | 9617338 |
| characterization of wild-type adeno-associated virus type 2-like particles generated during recombinant viral vector production and strategies for their elimination. | the psub201-paav/ad plasmid cotransfection system was developed to eliminate homologous recombination which leads to generation of the wild-type (wt) adeno-associated virus type 2 (aav) during recombinant vector production. the extent of contamination with wt aav has been documented to range between 0.01 and 10%. however, the precise mechanism of generation of the contaminating wt aav remains unclear. to characterize the wt aav genomes, recombinant viral stocks were used to infect human 293 cell ... | 1998 | 9621003 |
| adeno-associated virus vector-mediated transgene integration into neurons and other nondividing cell targets. | the site-specific integration of wild-type adeno-associated virus (wtaav) into the human genome is a very attractive feature for the development of aav-based gene therapy vectors. however, knowledge about integration of wtaav, as well as currently configured recombinant aav (raav) vectors, is limited. by using a modified alu-pcr technique to amplify and sequence the vector-cellular junctions, we provide the first direct evidence both in vitro and in vivo of raav-mediated transgene integration in ... | 1998 | 9621054 |
| site-specific integration of adeno-associated virus into an episome with the target locus via a deletion-substitution mechanism. | five site-specific adeno-associated virus integrants generated in a model system with an epstein-barr virus- based shuttle vector have been characterized. the results suggest a deletion-substitution mechanism of recombination. | 1998 | 9621089 |
| selective and rapid uptake of adeno-associated virus type 2 in brain. | recombinant adeno-associated virus (aav) vectors effectively transfer and express foreign genes in the brain. the transferred genes, however, are selectively expressed in neurons, and the cause of this specificity is not understood. to address this question, wild-type aav-2 capsids were covalently labeled with the fluorophore, cy3, and infused into the inferior colliculus or the hippocampus. using antibodies to identify neurons (neun), astrocytes (gfap), or oligodendrocytes (ox-42), clear neuron ... | 1998 | 9625257 |
| 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 |
| transduction of human trophoblastic cells by replication-deficient recombinant viral vectors. promoting cellular differentiation affects virus entry. | we investigated the transfer of the lacz reporter gene into human trophoblastic cells using herpes simplex virus and adeno-associated virus vectors. we used an established choriocarcinoma cell line (bewo cells) that can be induced to terminally differentiate after treatment with cyclic-amp. our results demonstrate that transduction of trophoblastic cells by the herpes simplex virus vector, hsv.cmvlac, and the adeno-associated virus vector, aav.cmvlac, is affected by cellular differentiation. tre ... | 1998 | 9626056 |
| discrimination between different types of human adeno-associated viruses in clinical samples by pcr. | persistent infection of human tissues with the helper virus-dependent parvovirus, adeno-associated virus (aav) was detected by polymerase chain reaction (pcr) using primer pairs detecting aav types 2, 3 or 5. in order to develop pcr protocols which discriminate between the different serotypes of aav, the dna of aav-5 was sequenced partially and compared with the published sequences of aav-2 and -3. type specific oligonucleotides and specific probes which allow the distinction between human aav t ... | 1998 | 9628217 |
| recombinant adeno-associated virus vector: use for transgene expression and anterograde tract tracing in the cns. | we used a recombinant adeno-associated virus vector (aav) to deliver a foreign gene, green fluorescent protein (gfp), into mature neurons in adult rat cns in vivo. microinjections of aav as small as 50 nl transduced hundreds of neurons at the injection site. there was virtually no retrograde transport as fewer than one neuron per brain was found distant from the injection site that exhibited gfp immunoreactivity. the gene product, gfp, filled the entire neuronal cytoplasmic compartment; gfp immu ... | 1998 | 9630611 |
| adeno-associated virus gene transfer into renal cells: potential for in vivo gene delivery. | the human parvovirus adeno-associated virus (aav), type 2, has a number of features that make it an attractive choice as a vector for gene delivery to the kidney. aav vectors permit long-term gene expression in vivo by integration into the host genome, have potential for site-specific integration on chromosome 19, do not express viral genes or generate a cellular immune response, and demonstrate enhancement of gene expression by chemotherapeutic agents that are approved for use in vivo. these pr ... | 1998 | 9639033 |
| prevention of dopaminergic neuron death by adeno-associated virus vector-mediated gdnf gene transfer in rat mesencephalic cells in vitro. | glial cell line-derived neurotrophic factor (gdnf) is known as a potent neurotrophic factor for dopaminergic neurons. since adeno-associated virus (aav) vector is a suitable vehicle for gene transfer into neurons, rat e14 mesencephalic cells were transduced with an aav vector expressing gdnf. when compared with mock transduction, a larger number of dopaminergic neurons survived in aav-gdnf-transduced cultures (234% and 325% of controls at 1 and 2 weeks, respectively; p < 0.01). furthermore, the ... | 1998 | 9665664 |
| detection of infectious adeno-associated virus particles in human cervical biopsies. | recently we reported that dna of the human oncogenic papillomaviruses (hpv) and the tumor suppressive human helper virus-dependent parvoviruses, adeno-associated viruses type 2 (aav-2), colocalize in cervical epithelium. to analyze whether infectious aav particles are present in cervical tissue, we examined cervical biopsies from 36 patients with hpv-related lesions (squamous intraepithelial lesions) for the presence of aav dna and of infectious aav. from each patient specimens from the lesion a ... | 1998 | 9683575 |
| adeno-associated virus-mediated gene transfer into human retinal pigment epithelium cells. | adeno-associated virus (aav) is emerging as a promising vector for gene therapy | 1998 | 9685031 |
| recent advances in skeletal-muscle-based gene therapy. | gene transfer into skeletal muscle holds promise for the treatment of a variety of serum protein deficiencies, muscular dystrophies, and chronic ischemic limb syndromes. the past two years have seen the development of new and improved vectors for programming recombinant gene expression in skeletal muscle. important advances include first, novel plasmid dna, adenovirus, and adeno-associated virus vectors that can be used to stably express therapeutic levels of recombinant proteins in the skeletal ... | 1998 | 9691001 |
| prospects for gene therapy in pediatric neurosurgery. | gene therapy represents a powerful tool for both the study and potential treatment of pediatric neurological diseases. the majority of strategies for brain gene therapy have focused upon the use of modified viruses as vehicles for efficient delivery of genes into cells of the central nervous system. retroviruses were originally the most popular vehicles for gene transfer outside the brain; however, these only function in actively dividing cells and have thus been limited to developmental neurobi ... | 1998 | 9693323 |
| gene therapy for genetic diseases. | gene therapy provides the potential to permanently cure selected genetic diseases. however, a major obstacle is the effective delivery of the normal gene to specific target sites of pathology and continuous expression at therapeutic levels. a variety of viral and non-viral vectors have been developed to deliver genes to various cells, tissues and organs by ex vivo and in vivo strategies. among the viral-based vectors, retroviruses, adenovirus, adeno-associated virus and herpes virus have been th ... | 1998 | 9695290 |
| packaging cells based on inducible gene amplification for the production of adeno-associated virus vectors. | although vectors based on adeno-associated virus (aav) offer several unique advantages, their usage has been hampered by the difficulties encountered in vector production. in this report, we describe a new aav packaging system based on inducible amplification of integrated helper and vector constructs containing the simian virus 40 (sv40) replication origin. the packaging and producer cell lines developed express sv40 t antigen under the control of the reverse tetracycline transactivator system, ... | 1998 | 9696794 |
| 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 |
| ribozyme rescue of photoreceptor cells in a transgenic rat model of autosomal dominant retinitis pigmentosa. | ribozymes, catalytic rna molecules that cleave a complementary mrna sequence, have potential as therapeutics for dominantly inherited disease. twelve percent of american patients with the blinding disease autosomal dominant retinitis pigmentosa (adrp) carry a substitution of histidine for proline at codon 23 (p23h) in their rhodopsin gene, resulting in photoreceptor cell death from the synthesis of the abnormal gene product. ribozymes can discriminate and catalyze the in vitro destruction of p23 ... | 1998 | 9701253 |
| regulation of gene expression in vivo following transduction by two separate raav vectors. | control of gene expression is important to gene therapy for purposes of both dosing and safety. in vivo regulation of gene expression was demonstrated following co-injection of two separate recombinant adeno-associated virus vectors, one encoding an inducible murine erythropoietin transgene and the other a transcriptional activator, directly into the skeletal muscle of adult immunocompetent mice. transcription was controlled by systemic administration or withdrawal of tetracycline over an 18 wee ... | 1998 | 9702775 |
| 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 |
| efficient expression of cftr function with adeno-associated virus vectors that carry shortened cftr genes. | adeno-associated virus (aav)-based vectors have been shown to be effective in transferring the cystic fibrosis gene (cftr) into airway epithelial cells in animal models and in patients. however, the level of cftr gene expression has been low because the vector cannot accommodate the cftr gene together with a promoter. in this study, we described a strategy to reduce the size of the cftr cdna to allow the incorporation of an effective promoter with the cftr gene into aav vectors. we engineered an ... | 1998 | 9707617 |
| efficient gene transfer into cardiac myocytes using adeno-associated virus (aav) vectors. | adeno-associated virus (aav) vectors, derived from a non-pathogenic parvovirus, are considered to be an appropriate gene transfer vehicle for both dividing and non-dividing cells. in the present study, we investigated whether the rat heart could be efficiently transduced with aav vectors. rat cardiac myocytes (cm) were infected with aav-lacz vector containing beta-galactosidase (beta-gal) gene in vitro, and the expression of beta-gal in cm was evaluated by x-gal staining and beta-gal elisa. with ... | 1998 | 9710802 |
| construction of mdr1 adeno-associated virus vectors for gene therapy. | 1998 | 9711581 | |
| adeno-associated virus vectors for gene therapy of cystic fibrosis. | 1998 | 9711594 | |
| control of erythropoietin delivery by doxycycline in mice after intramuscular injection of adeno-associated vector. | we reported previously that controlled expression of a foreign gene in response to tetracycline derivative can be accomplished in mice by the autologous transplantation of retrovirus-modified muscle cells. although regulated systemic delivery of therapeutic proteins from engineered tissues has potential clinical application, the transplantation of muscle cells is not currently feasible in humans. several studies have shown that a single injection of adeno-associated virus (aav) vectors into mous ... | 1998 | 9716577 |
| adeno-associated virus rep78 protein interacts with protein kinase a and its homolog prkx and inhibits creb-dependent transcriptional activation. | adeno-associated virus (aav) is a human parvovirus of the genus dependovirus. aav replication is largely restricted to cells which are coinfected with a helper virus. in the absence of a helper virus, the aav genome can integrate into a specific chromosomal site where it remains latent until reactivated by superinfection of the host cell with an appropriate helper virus. replication functions of aav have been mapped to the rep68 and rep78 gene products. rep proteins demonstrate dna binding, endo ... | 1998 | 9733829 |
| an adenovirus type 5 mutant with the preterminal protein gene deleted efficiently provides helper functions for the production of recombinant adeno-associated virus. | production of recombinant adeno-associated virus (raav) requires helper functions that have routinely been provided by infection of the producer cells with adenovirus. complete removal and/or inactivation of progeny adenovirus, present in such raav preparations, presents significant difficulty. here, we report that an adenovirus type 5 (ad5) mutant with the preterminal protein (ptp) gene deleted can provide helper function for the growth of raav. at high multiplicity, ad5dl308deltaptp was as eff ... | 1998 | 9733887 |
| solution structure of a na cation stabilized dna quadruplex containing g.g.g.g and g.c.g.c tetrads formed by g-g-g-c repeats observed in adeno-associated viral dna. | we have applied nmr and molecular dynamics computations including intensity based refinement to define the structure of the d(g-g-g-c-t4-g-g-g-c) dodecanucleotide in 100 mm nacl solution. the g-g-g-c sequence is of interest since it has been found as tandem repeats in the dna sequence of human chromosome 19. the same g-g-g-c sequence is also seen as islands in adeno-associated virus, a human parvovirus, which is unique amongst eukaryotic dna viruses in its ability to integrate site-specifically ... | 1998 | 9737926 |
| inhibition of prkx, a novel protein kinase, and the cyclic amp-dependent protein kinase pka by the regulatory proteins of adeno-associated virus type 2. | adeno-associated virus encodes four nonstructural proteins, which are known as rep78, rep68, rep52, and rep40. expression of these nonstructural proteins affects cell growth and gene expression through processes that have not yet been characterized. using a yeast two-hybrid screen, we have demonstrated that a stable interaction occurs between the viral proteins rep78 and rep52 and the putative protein kinase prkx, which is encoded on the x chromosome. the stability and specificity of the rep-prk ... | 1998 | 9742109 |
| in vivo expression of therapeutic human genes for dopamine production in the caudates of mptp-treated monkeys using an aav vector. | an adeno-associated virus (aav) vector, expressing genes for human tyrosine hydroxylase (th) and aromatic amino acid decarboxylase (aadc), demonstrated significantly increased production of dopamine in 293 (human embryonic kidney) cells. this bicistronic vector was used to transduce striatal cells of six asymptomatic but dopamine-depleted monkeys which had been treated with the neurotoxin mptp. striatal cells were immunoreactive for the vector-encoded th after stereotactic injection for periods ... | 1998 | 9747462 |
| transcription-positive cofactor 4 enhances rescue of adeno-associated virus genome from an infectious clone. | while rep proteins are required for adeno-associated virus (aav) replication, little is known about cellular proteins that interact with rep. we demonstrate here that transcription-positive cofactor 4 (pc4, p15) fused to gal4-activating domain interacted with both aav-2 and aav-3 rep proteins fused to gal4 dna-binding domain, leading to reporter activation in the yeast two-hybrid system. in addition to its coactivating function, pc4 recently has been shown to be involved in replication of simian ... | 1998 | 9747724 |
| circular intermediates of recombinant adeno-associated virus have defined structural characteristics responsible for long-term episomal persistence in muscle tissue. | adeno-associated viral (aav) vectors have demonstrated great utility for long-term gene expression in muscle tissue. however, the mechanisms by which recombinant aav (raav) genomes persist in muscle tissue remain unclear. using a recombinant shuttle vector, we have demonstrated that circularized raav intermediates impart episomal persistence to raav genomes in muscle tissue. the majority of circular intermediates had a consistent head-to-tail configuration consisting of monomer genomes which slo ... | 1998 | 9765395 |
| factors influencing adeno-associated virus-mediated gene transfer to human cystic fibrosis airway epithelial cells: comparison with adenovirus vectors. | adeno-associated virus (aav) vectors appear promising for use in gene therapy in cystic fibrosis (cf) patients, yet many features of aav-mediated gene transfer to airway epithelial cells are not well understood. we compared the transduction efficiencies of aav vectors and adenovirus (ad) vectors in immortalized cell lines from cf patients and in nasal epithelial primary cultures from normal humans and cf patients. similar dose-dependent relationships between the vector multiplicities of infectio ... | 1998 | 9765435 |
| peroral gene therapy of lactose intolerance using an adeno-associated virus vector. | gene therapy is usually reserved for severe and medically refractory disorders because of the toxicity, potential long-term risks and invasiveness of most gene transfer protocols. here we show that an orally administered adeno-associated viral vector leads to persistent expression of a beta-galactosidase transgene in both gut epithelial and lamina propria cells, and that this approach results in long-term phenotypic recovery in an animal model of lactose intolerance. a gene 'pill' associated wit ... | 1998 | 9771745 |
| adeno-associated virus vector containing the herpes simplex virus thymidine kinase gene causes complete regression of intracerebrally implanted human gliomas in mice, in conjunction with ganciclovir administration. | adeno-associated virus (aav) has attracted considerable interest as a potential vector for gene therapy because of its wide host range, high transduction efficiency, and lack of cytopathogenicity. in this experiment, we evaluated the efficacy of aav vector containing the herpes simplex virus thymidine kinase (hsv-tk) gene on human gliomas transplanted into the brain of nude mice. complete regression of the tumors was observed after multiple aav-tk injections followed by intraperitoneal ganciclov ... | 1998 | 9510479 |
| reduced mk801 binding in neocortical neurons after aav-mediated transfections with nmda-r1 antisense cdna. | two adeno-associated virus (aav)-derived plasmids were constructed with portions of the n-methyl-d-aspartic acid-r1 (nmda-r1) receptor subunit downstream from the aav p40-(pjdt95dlk-ar1) or cytomegalovirus (cmv) promoter (ptruf3-ar1) in an antisense orientation. each plasmid drove expression of antisense nmda-r1 in primary rat neocortical neuronal cultures 4 days after transfection as detected by reverse transcriptase-polymerase chain reaction (rt-pcr). transfection with ptruf3-ar1 (2x4 microgra ... | 1998 | 9518673 |
| cellular proteins required for adeno-associated virus dna replication in the absence of adenovirus coinfection. | we previously reported the development of an in vitro adeno-associated virus (aav) dna replication system. the system required one of the p5 rep proteins encoded by aav (either rep78 or rep68) and a crude adenovirus (ad)-infected hela cell cytoplasmic extract to catalyze origin of replication-dependent aav dna replication. however, in addition to fully permissive dna replication, which occurs in the presence of ad, aav is also capable of partially permissive dna replication in the absence of the ... | 1998 | 9525597 |
| in vitro packaging of adeno-associated virus dna. | we have developed an in vitro procedure for packaging of recombinant adeno-associated virus (aav). by using aav replicative-form dna as the substrate, it is possible to synthesize an infectious aav particle in vitro that can be used to transfer a marker gene to mammalian cells. the packaging procedure requires the presence of both the aav rep and capsid proteins. two kinds of in vitro products can be formed which facilitate dna transfer. both are resistant to heat and have a density in cesium ch ... | 1998 | 9525651 |