| viral vectors for modulating gene expression in neurons. | the inability to reliably express foreign proteins in postmitotic neurons has hampered numerous studies in the field of neurobiology. within the past several years, a number of viral vectors that overcome this problem under controlled conditions in vitro have been developed. in particular, recombinant adenoviruses have proved to be efficient, non-cytotoxic vectors for manipulating neurons in dissociated and organotypic cultures, when used at low viral titres. in contrast, vectors derived from he ... | 1996 | 8937820 |
| gene therapy against an experimental glioma using adeno-associated virus vectors. | the efficacy of gene therapy for glioma was examined using adeno-associated virus (aav)-based vectors to deliver genes to experimental tumors in mice. stereotactic injection of 2 x 10(5) u-251sp human glioma cells into the brains of nude mice produced tumors of 19.06 +/- 1.79 mm2 17 days after injection. employing a high titer preparation of aav vector containing the gene for beta-galactosidase (aav-lacz), dose-dependent transduction of u-251sp cells was seen in vitro. when 1.6 x 10(10) aav-lacz ... | 1996 | 8940635 |
| role of the terminal repeat gagc trimer, the major rep78 binding site, in adeno-associated virus dna replication. | the adeno-associated virus (aav) terminal repeats (tr) are cis required, and the aav encoded rep78 protein is trans required, for aav dna replication. the rep78 protein recognizes and interacts with at least three regions within the tr dna. the major binding site, with the highest affinity for rep78 binding, is within the tr stem (nt 36-16) and includes the 'core' gagc trimer (gagc3, nt 33-22; fig. 2) sequence. in this study mutations were made within the gagc trimer and these mutants assayed fo ... | 1996 | 8941722 |
| cocrystal structure of yy1 bound to the adeno-associated virus p5 initiator. | ying-yang 1 protein (yy1) supports specific, unidirectional initiation of messenger rna production by rna polymerase ii from two adjacent start sites in the adeno-associated virus p5 promoter, a process which is independent of the tata box-binding protein (tbp). the 2.5-a resolution yy1-initiator element cocrystal structure reveals four zinc fingers recognizing a yy1-binding consensus sequence. upstream of the transcription start sites protein-dna contacts involve both strands and downstream the ... | 1996 | 8942976 |
| gene delivery to skeletal muscle results in sustained expression and systemic delivery of a therapeutic protein. | somatic gene therapy has been proposed as a means to achieve systemic delivery of therapeutic proteins. however, there is limited evidence that current methods of gene delivery can practically achieve this goal. in this study, we demonstrate that, following a single intramuscular administration of a recombinant adeno-associated virus (raav) vector containing the beta-galactosidase (aav-lacz) gene into adult balb/c mice, protein expression was detected in myofibers for at least 32 weeks. a single ... | 1996 | 8943064 |
| long-term gene transfer in porcine myocardium after coronary infusion of an adeno-associated virus vector. | viral vector-mediated gene transfer into the heart represents a potentially powerful tool for studying both cardiac physiology as well as gene therapy of cardiac disease. we report here the use of a defective viral vector, which expresses no viral gene products, for gene transfer into the mammalian heart. previous studies have used recombinant viral vectors, which retained viral genes and yielded mostly short-term expression, often with significant inflammation. | 1996 | 8957370 |
| 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 |
| a stable cell line carrying adenovirus-inducible rep and cap genes allows for infectivity titration of adeno-associated virus vectors. | adeno-associated virus (aav) vectors are being developed for in vivo and ex vivo gene transfer to human cells. at present, widespread usage of aav vectors is limited primarily by difficulties in generating recombinant virions on a scale sufficient for in-depth preclinical and clinical trials. however, recent work in several laboratories suggests that this technical obstacle should be overcome in the near future. as a result, it can be anticipated that the interest in aav vectors will expand, thu ... | 1996 | 8986439 |
| 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 |
| stable transduction of recombinant adeno-associated virus into hematopoietic stem cells from normal and sickle cell patients. | stable introduction of genes into human hematopoietic stem cells with self-renewing potential is a necessary requirement for gene therapy strategies. we have developed an adeno-associated virus (aav) vector and a partial packaging cell line that produces recombinant aav at a titer of 10(8) transducing particles per milliliter. a high-titer viral stock containing the cmv/lacz gene was used to transfer lacz sequences into cd34+ lin-thy+ hematopoietic stem cells purified from normal and homozygous ... | 1996 | 9078351 |
| characterization of recombinant adeno-associated virus-2 as a vehicle for gene delivery and expression into vascular cells. | we have used wild-type and recombinant adeno-associated virus-2 (aav) to study transduction, replication efficiencies, functional protein expression, and gene delivery to vascular cells in vitro and in vivo. | 1997 | 9084579 |
| detection of adeno-associated virus type 2 in sorted human bone marrow progenitor cells. | wild-type adeno-associated virus (wtaav) is a helper-dependent human parvovirus which has the ability to integrate into the genome of a wide variety of human cells, including those of the hemopoietic lineages. recombinant adeno-associated virus (raav) is becoming a good candidate for virally mediated gene therapy. raav is likely to be a safe vector in clinical gene transfer, as it has never been associated with any disease despite previous studies showing that up to 70% of adults are seropositiv ... | 1997 | 9091303 |
| evaluation of beta-globin gene therapy constructs in single copy transgenic mice. | effective gene therapy constructs based on retrovirus or adeno-associated virus vectors will require regulatory elements that direct expression of genes transduced at single copy. most beta-globin constructs designed for therapy of beta-thalassemias are regulated by the 5'hs2 component of the locus control region (lcr). here we show that a human beta-globin gene flanked by two small 5'hs2 core elements or flanked by a 5'hs3 (footprints 1-3) core and a 5'hs2 core are not reproducibly expressed in ... | 1997 | 9092642 |
| potent inhibition of human immunodeficiency virus type 1 in primary t cells and alveolar macrophages by a combination anti-rev strategy delivered in an adeno-associated virus vector. | the rate of viral replication appears to play a pivotal role in human immunodeficiency virus type 1 (hiv-1) pathogenesis and disease progression as it outstrips the capacity of the immune system to respond. important cellular sites for hiv-1 production include t lymphocytes and tissue macrophages. antiviral strategies, including newer treatment modalities such as gene therapy of hiv-1-susceptible cell populations, must be capable of engendering durable inhibitory effects to hiv-1 replication in ... | 1997 | 9094685 |
| active immunization with tumor cells transduced by a novel aav plasmid-based gene delivery system. | ex vivo genetically engineered cytokine-secreting tumor cell vaccines have been shown to prevent metastatic disease in animal models of lung and breast cancer. because of the inefficiency of existing modes of gene delivery in transducing primary human tumor cells, it has been difficult to clinically apply this strategy. in this study, liposome-mediated delivery of an adeno-associated virus (aav)-based plasmid containing the sequence for murine gamma-interferon (gamma-ifn) (pmp6a-mifn-gamma) was ... | 1997 | 9101411 |
| recombinant adeno-associated viral vectors mediate long-term transgene expression in muscle. | gene transfer to muscle holds overt promise for the treatment of inherited myopathies, lysosomal storage disorders, and serum protein deficiencies. in addition, muscle could provide a reservoir for delivery of therapeutic molecules like blood clotting factors, erythropoietin, or insulin. to date, successful gene transfer to muscle has been limited by the inefficiency of the vector delivery systems and the transient nature of gene expression. in this paper, we show that a vector based on recombin ... | 1997 | 9113506 |
| adeno-associated virus vectors for vascular gene delivery. | a variety of delivery systems have been used to genetically modify vascular endothelial cells and smooth muscle cells (smcs), but currently available systems suffer from either inefficient in vivo gene transfer, transient episomal vector expression, or significant immune responses and inflammation. in the present study, we evaluated an alternate vector system, recombinant adeno-associated virus (raav) for transduction of vascular cells in culture and in vivo. primary cultures of rabbit, monkey, ... | 1997 | 9118480 |
| characterization of the rep78/adeno-associated virus complex. | adeno-associated virus (aav) replication proteins rep78 and rep68 play major roles in the life cycle of aav. we have recently provided in vivo evidence for the existence of a covalent association between rep78 and virion single-stranded (ss) aav dna (k. m. r. prasad and j.p. trempe(1995) virology 214, 360-370). in this work we have further characterized the rep78 protein-aav dna covalent linkage. exonuclease and primer extension analyses revealed that in the majority of isolated ssdna, rep78 pro ... | 1997 | 9123860 |
| defective viral vectors as agents for gene transfer in the nervous system. | viral vectors have attracted great interest as vehicles for gene therapy. due to concerns regarding continued viral gene expression in several systems, new approaches have been sought for gene transfer in the nervous system. this article reviews the general concepts and basic biology of defective viral vectors. these are vectors which can package into a viral coat but contain no viral genes, thereby allowing efficient gene transfer in the absence of viral gene expression in target cells. the def ... | 1997 | 9125381 |
| gene therapeutic strategies for neuroprotection: implications for parkinson's disease. | gene transfer methodologies are being explored as strategies to restore and preserve neuronal function in parkinson's disease. this technology represents a new therapeutic modality, holding promise for continuous and localized delivery, of neuroprotective molecules. two primary approaches for gene transfer have emerged: in vivo and ex vivo. recent advances in the construction and characterization of gene transfer vectors have generated more efficient vehicles to deliver and express candidate the ... | 1997 | 9126153 |
| gene transfer by adeno-associated virus vectors into the central nervous system. | adeno-associated virus (aav) vectors are derived from a nonpathogenic and defective human parvovirus. although currently unable to display the integration specificity featured by its wild-type parent, the recombinant aav (raav) system has continued to attract enormous interest primarily due to its unique features such as safety, high titers, broad host range, transduction of quiescent cells, and vector integration. recently, raav-mediated in vivo gene transfers have demonstrated efficient long-t ... | 1997 | 9126160 |
| structural and functional heterogeneity of integrated recombinant aav genomes. | adeno-associated virus (aav) has emerged as a promising vector for gene therapy because of its ability to generate high titer recombinant stocks and the potential for site specific integration. however, much of the current knowledge regarding the transduction and integration biology of this virus is based on studies evaluating wild type aav or recombinant aav which was unknowingly contaminated with wild type virus. given the fact that recombinant aav is replication incompetent, by virtue of dele ... | 1997 | 9140193 |
| the packaging capacity of adeno-associated virus (aav) and the potential for wild-type-plus aav gene therapy vectors. | because of its ability to integrate chromosomally and its non-pathogenic nature, adeno-associated virus (aav) has significant potential as a human gene therapy vector. here we investigate the maximum amount of dna which can be inserted into the aav genome and still allow efficient packaging into an infectious virus particle. altered wild-type aav genomes were constructed with inserts, which increased in size by 100 bp, ligated at map unit 96. these large wild-type-plus genomes were able to repli ... | 1997 | 9141485 |
| the adeno-associated virus type 2 p40 promoter requires a proximal sp1 interaction and a p19 carg-like element to facilitate rep transactivation. | we have identified the sequence elements that are required for adeno-associated virus type 2 p40 promoter activity. mutation of specific promoter elements showed that two sp1 sites at approximately -50 (sp1-50) and -70 (ggt-70) bp upstream of the start of the p40 messages were necessary for maximal promoter activity. as expected, the tata site at -30 was also essential. in vitro dna binding experiments confirmed that the sp1-50 and ggt-70 sites were bound by sp1 or sp1-like proteins. two other t ... | 1997 | 9151818 |
| the rep78 gene product of adeno-associated virus (aav) self-associates to form a hexameric complex in the presence of aav ori sequences. | the rep78 and rep68 proteins of adeno-associated virus (aav) are replication initiator proteins that bind the viral replicative-form origin of replication, nick the origin in a site- and strand-specific fashion, and mediate vectorial unwinding of the dna duplex via an atp-dependent helicase activity, thus initiating a strand displacement mechanism of viral dna replication. genetic and biochemical studies have identified rep mutants that demonstrate a trans-dominant negative phenotype in vitro an ... | 1997 | 9151837 |
| stable gene transfer and expression of human blood coagulation factor ix after intramuscular injection of recombinant adeno-associated virus. | we sought to determine whether intramuscular injection of a recombinant adeno-associated virus (raav) vector expressing human factor ix (hf.ix) could direct expression of therapeutic levels of the transgene in experimental animals. high titer (10(12)-10(13) vector genomes/ml) raav expressing hf.ix was prepared, purified, and injected into hindlimb muscles of c57bl/6 mice and rag 1 mice. in the immunocompetent c57bl/6 mice, immunofluorescence staining of muscle harvested 3 months after injection ... | 1997 | 9159155 |
| adeno-associated virus 2-mediated gene transfer in vivo: organ-tropism and expression of transduced sequences in mice. | adeno-associated virus 2 (aav), a non-pathogenic human parvovirus, is gaining attention as a vector for its potential use in human gene therapy. however, few studies have examined the safety and the efficacy of this vector system in vivo. we report here that recombinant aav vectors, when directly injected intravenously in mice, accumulated predominantly in liver cells, suggesting that aav may possess in vivo organ-tropism for liver. the transduced lacz reporter gene was expressed in hepatocytes ... | 1997 | 9185868 |
| adeno-associated virus (aav) vector antisense gene transfer in vivo decreases gaba(a) alpha1 containing receptors and increases inferior collicular seizure sensitivity. | in the inferior colliculus, adeno-associated virus (aav) vectors are capable of gene transfer and stable, long-term expression, but it remained to be shown if this in vivo gene transfer could alter focal seizure sensitivity in the inferior colliculus. because gaba receptors directly modulate inferior collicular seizures, aav vectors were constructed with a cytomegalovirus (cmv) promoter and a truncated, human gaba(a) alpha1 cdna in both the sense and antisense orientations. seven days after coll ... | 1997 | 9187316 |
| role for highly regulated rep gene expression in adeno-associated virus vector production. | recent success achieving long-term in vivo gene transfer without a significant immune response by using adeno-associated virus (aav) vectors (x. xiao, j. li, and r. j. samulski, j. virol. 70:8098-8108, 1996) has encouraged further development of this vector for human gene therapy. currently, studies focus on the generation of high-titer vectors by using the two-plasmid helper-vector system in adenovirus (ad)-infected cells. to examine the effects of the aav replication (rep) genes on recombinant ... | 1997 | 9188591 |
| adeno-associated virus type 2-mediated transfer of ecotropic retrovirus receptor cdna allows ecotropic retroviral transduction of established and primary human cells. | the cellular receptors that mediate binding and internalization of retroviruses have recently been identified. the concentration and accessibility of these receptors are critical determinants in accomplishing successful gene transfer with retrovirus-based vectors. murine retroviruses containing ecotropic glycoproteins do not infect human cells since human cells do not express the receptor that binds the ecotropic glycoproteins. to enable human cells to become permissive for ecotropic retrovirus- ... | 1997 | 9188645 |
| construction and biological characterization of an interleukin-12 fusion protein (flexi-12): delivery to acute myeloid leukemic blasts using adeno-associated virus. | interleukin-12 (il-12) is a cytokine that exhibits pleiotropic effects on lymphocytes and natural killer cells and has been shown to have promise for the immunotherapy of cancer. the combination of the immune costimulatory molecule b7.1 and il-12 has been shown to be synergistic for t cell activation. by transfecting tumor cells with both il-12 and b7.1 cdnas, it may be possible to use these modified targets as vaccines. a major obstacle in designing a vector to deliver these genes results from ... | 1997 | 9189770 |
| phosphorylation of the adeno-associated virus replication proteins. | the adeno-associated virus (aav) replication proteins rep78 and rep68 regulate viral gene expression and dna amplification. their effects on both processes suggest that they play roles in all phases of the virus life cycle. we have investigated rep protein phosphorylation to determine if this modification might alter rep function. all four rep proteins were found to be phosphorylated in aav and adenovirus co-infected cell cultures, and rep proteins contained phospho-serine whereas no phospho-thr ... | 1997 | 9191846 |
| interaction of human papillomavirus type 16 and adeno-associated virus type 2 co-infecting human cervical epithelium. | recently, we hypothesized that the tumour-suppressive, human helper-virus-dependent, adeno-associated parvoviruses (aav) may interfere with transforming functions of human papillomaviruses (hpv) in the development of cervical carcinoma. here, we demonstrate that in cervical epithelium containing papillomavirus dna, aav dna can be detected in a replication-competent form and that aav proteins are expressed. in cultured cells containing integrated aav-2 dna, transfection of hpv-16 dna induced resc ... | 1997 | 9191942 |
| assembly of adeno-associated virus type 2 capsids in vitro. | capsid proteins vp1, vp2 and vp3 of adeno-associated virus type 2 (aav-2) were separately expressed by recombinant baculoviruses, purified under denaturing conditions and renatured in the presence of 0.5 m arginine, followed by dialysis against buffers of physiological ionic strength. at a protein concentration of 0.05 mg/ml, the three capsid proteins predominantly formed monomers and, to a lesser extent, oligomers, as determined by sedimentation analysis. oligomerization increased at higher pro ... | 1997 | 9191943 |
| efficient photoreceptor-targeted gene expression in vivo by recombinant adeno-associated virus. | we describe a general approach for achieving efficient and cell type-specific expression of exogenous genes in photoreceptor cells of the mammalian retina. recombinant adeno-associated virus (raav) vectors were used to transfer the bacterial lacz gene or a synthetic green fluorescent protein gene (gfp) to mouse or rat retinas after injection into the subretinal space. using a proximal murine rod opsin promoter (+86 to -385) to drive expression, reporter gene product was found exclusively in phot ... | 1997 | 9192666 |
| construction of recombinant adeno-associated virus vector containing the rat preproinsulin ii gene. | we have investigated a possible delivery system for the rat preproinsulin ii gene (ri2) utilising a recombinant adeno-associated virus (raav) vector system, with the long-term goal of engineering stably infected insulin-producing cell lines. the raav vector was chosen because it is a safe and nonpathogenic method for gene transfer. the plasmid pbc12bi (atcc) was purified and digested with restriction enzymes sepi and stui to release a fragment containing the rous sarcoma virus long terminal repe ... | 1997 | 9202669 |
| persistent and therapeutic concentrations of human factor ix in mice after hepatic gene transfer of recombinant aav vectors. | haemophilia b, or factor ix deficiency, is a x-linked recessive disorder that occurs in about one in 25,000 males, and severely affected people are at risk for spontaneous bleeding into numerous organs. bleeding can be life-threatening or lead to chronic disabilities with haemophilic arthropathy. the severity of the bleeding tendency varies among patients and is related to the concentration of functional plasma factor ix. patients with 5-30% of the normal factor ix have mild haemophilia that may ... | 1997 | 9207793 |
| adeno-associated viral vector gene transfer into leptomeningeal xenografts. | leptomeningeal carcinomatosis is a painful and debilitating complication of cancer. indwelling reservoirs provide continuous assess to the subarachnoid space, making leptomeningeal cancer potentially amenable to gene therapy. adeno-associated virus (aav) is a defective virus not associated with any human disease. we used an aav vector to transduce medulloblastoma (daoy) cells in a nude rat model of leptomeningeal disease. after intraventricular injection of vector carrying the bacterial lacz gen ... | 1997 | 9210060 |
| in vivo gene therapy with adeno-associated virus vectors for cystic fibrosis. | | 1997 | 9217924 |
| transduction by adeno-associated virus vectors in the rabbit airway: efficiency, persistence, and readministration. | the ability of recombinant adeno-associated virus (aav) vectors to integrate into the host genome and to transduce nondividing cells makes them attractive as vehicles for gene delivery. in this study, we assessed the ability of several aav vectors to transduce airway cells in rabbits by measuring marker gene expression. aav vectors that transferred either a beta-galactosidase (beta-gal) or a human placental alkaline phosphatase (ap) gene were delivered to one lobe of the rabbit lung by use of a ... | 1997 | 9223483 |
| infection of primary cells by adeno-associated virus type 2 results in a modulation of cell cycle-regulating proteins. | it has been demonstrated that infection of primary human cells with adeno-associated viruses (aav) leads to a decrease in cellular proliferation and to growth arrest. we analyzed the molecular basis of this phenomenon and observed that infection with aav type 2 (aav2) had an effect on several factors engaged in the control of the mammalian cell cycle. in particular, all of the prb family members, prb, p107, and p130, which are involved in g1 cell cycle checkpoint control, were affected. after in ... | 1997 | 9223493 |
| transduction of the human immunodeficiency virus type 1 promoter into human chromosomal dna by adeno-associated virus: effects on promoter activity. | transcription of the human immunodeficiency virus type 1 (hiv-1) genome takes place after integration of the provirus into human chromosomal dna. hiv transcription is known to be modulated by viral and cellular factors but the influence of flanking chromosomal sequences on proviral gene expression has not been well defined. to investigate the activity of the integrated hiv promoter, we exploited the ability of recombinant adeno-associated virus (aav-2) to transfer and stably integrate genes into ... | 1997 | 9234945 |
| potential use of herpes simplex virus (hsv) vectors for gene therapy of neurological disorders. | advances in molecular biology and virus genetics have allowed the possibility of gene therapy using viral vectors for a variety of neurological diseases in which the genetic or biochemical basis is understood. a number of such vectors have now been constructed, including those derived from herpes simplex virus (hsv), adenovirus, retrovirus and adeno-associated virus, and used in preliminary in vitro experiments and in animal models. it is possible to package a foreign gene into such a vector whi ... | 1997 | 9236634 |
| human immunodeficiency virus-1 proviral gene disruption by targeted gene therapy: a hypothetical technique for the elimination of provirus from the infected cells. | a hypothetical technique is proposed for the elimination of all the integrated human immunodeficiency virus-1 provirus from infected cells, based on the developing technology of selective gene excision through homologous recombination. in this technique, a recombinant retroviral packaging cell-line which would produce integrase-rep78 chimeric protein would be constructed. replication defective viral stocks would be made from this system which would have recombinant integrase-rep78 protein packag ... | 1997 | 9247905 |
| highly efficient and sustained gene transfer in adult neurons with a lentivirus vector. | the identification of monogenic and complex genes responsible for neurological disorders requires new approaches for delivering therapeutic protein genes to significant numbers of cells in the central nervous system. a lentivirus-based vector capable of infecting dividing and quiescent cells was investigated in vivo by injecting highly concentrated viral vector stock into the striatum and hippocampus of adult rats. control brains were injected with a moloney murine leukemia virus, adenovirus, or ... | 1997 | 9261386 |
| identification and elimination of replication-competent adeno-associated virus (aav) that can arise by nonhomologous recombination during aav vector production. | adeno-associated virus (aav) vector preparations are often contaminated with variable amounts of replication-competent aav (rcaav), which may influence the behavior of these vectors both in cultured cells and in animals. a packaging plasmid/vector plasmid system containing no significant homology and lacking the wild-type aav p5 promoter was constructed to eliminate the production of wild-type aav by recombination. still, rcaav was detected in vector produced by cotransfection of these plasmids ... | 1997 | 9261406 |
| cloning of adeno-associated virus type 4 (aav4) and generation of recombinant aav4 particles. | we have cloned and characterized the full-length genome of adeno-associated virus type 4 (aav4). the genome of aav4 is 4,767 nucleotides in length and contains an expanded p5 promoter region compared to aav2 and aav3. within the inverted terminal repeat (itr), several base changes were identified with respect to aav2. however, these changes did not affect the ability of this region to fold into a hairpin structure. within the itr, the terminal resolution site and rep binding sites were conserved ... | 1997 | 9261407 |
| mutational analysis of the adeno-associated virus type 2 rep68 protein helicase motifs. | the adeno-associated virus type 2 (aav) rep78 and rep68 proteins are required for viral replication. these proteins are encoded by unspliced and spliced transcripts, respectively, from the p5 promoter of aav and therefore have overlapping amino acid sequences. the rep78 and rep68 proteins share a variety of activities including endonuclease, helicase, and atpase activities and the ability to bind aav hairpin dna. the part of the amino acid sequence which is identical in rep78 and rep68 contains ... | 1997 | 9261429 |
| human genital tissues containing dna of adeno-associated virus lack dna sequences of the helper viruses adenovirus, herpes simplex virus or cytomegalovirus but frequently contain human papillomavirus dna. | the detection of dna of the helper virus-dependent adeno-associated virus type 2 (aav-2) in biopsies of material from spontaneous abortion and in tissue samples from the uterus raises the question of whether sequences of known helper viruses can be detected simultaneously within the same specimen despite the lack of histological evidence for the presence of lytic viruses. therefore, we performed pcr analyses with primers detecting dna sequences of viruses (adenovirus, herpes simplex virus and hu ... | 1997 | 9266994 |
| cftr gene transduction in neonatal rabbits using an adeno-associated virus (aav) vector. | patients with cystic fibrosis develop lung disease after birth, therefore cftr gene replacement therapy should be most efficacious in the neonatal period prior to the onset of pulmonary damage. an adeno-associated virus (aav) vector, sa306 (flotte tr et al proc natl acad sci usa 1993; 90: 10613-10617), which contains the aav inverted terminal repeats flanking the human cftr cdna linked to an amino-terminal epitope tag, was used to transduce a human cftr fusion protein into neonatal new zealand w ... | 1997 | 9274714 |
| [gene transfer into the patellar tendon of rabbits: a preliminary study of locoregional expression of growth factors]. | growth factors have the potential to enhance native repair responses in ligamentous and meniscal lesions. however, methods for applying these cytokines to sites of injury for extended periods are lacking. we suggest that local transfer of genes which encode the relevant healing factors merits investigation as a potential solution to this problem. in the present study different viral vectors and liposomes were evaluated for their ability to deliver genes to cells of ligamentous and meniscal origi ... | 1997 | 9281228 |
| cancer gene therapy using a novel adeno-associated virus vector expressing human wild-type p53. | previous studies have indicated that transfer of wild-type (wt) p53 cdna into cancer cells can suppress the tumor phenotype in vitro and in vivo. in this study we examined the effects of wt p53 transduction in the human cancer cell line h-358 (that bears a homozygous deletion of p53) using a novel recombinant adeno-associated viral vector engineered to express wt p53 (raavp53). western blot analysis demonstrated the expression of wt p53 in h-358 cells following infection with raavp53. furthermor ... | 1997 | 9282168 |
| rescue of photoreceptor function by aav-mediated gene transfer in a mouse model of inherited retinal degeneration. | knowledge of the mutations leading to inherited retinal degenerations provides a foundation for the development of somatic gene therapy in which potentially corrective genes are transferred to the target photoreceptor cells. towards this end, we have evaluated the efficacy of a recombinant adeno-associated virus (aav) vector to deliver and express the correct form of the cgmp phosphodiesterase-beta (pde-beta) gene in the retinas of rd mice, which suffer rapid retinal degeneration due to recessiv ... | 1997 | 9282169 |
| gene transfer of the costimulatory molecules b7-1 and b7-2 into human multiple myeloma cells by recombinant adeno-associated virus enhances the cytolytic t cell response. | gene transfer of the costimulatory molecules b7-1 and b7-2 induces a potent antitumor immune response in a variety of tumor models. b cell neoplasms including multiple myeloma (mm) often show little or no expression of b7 antigens; they are therefore a potential target for this approach. to increase the expression of human b7 genes in mm cells, both genes and the neomycin phosphotransferase gene were packaged into recombinant adeno-associated virus vectors (raav). the resulting recombinant virus ... | 1997 | 9282174 |
| [gene transfer using adeno-associated virus (aav) vectors]. | adeno-associated virus (aav) is a non-pathogenic, replication defective parvovirus. in the absence of helper adenovirus, aav stably integrates into a defined region of human chromosome 19. because of these unique properties, recombinant aav is considered to be an attractive vector for human gene therapy. it has been demonstrated that aav vectors are capable of efficient transduction of various types of cells including hematopoietic cells and post mitotic neuronal cells. we have recently develope ... | 1997 | 9284438 |
| evaluation of recombinant adeno-associated virus as a gene transfer vector for the retina. | to evaluate recombinant adeno-associated virus (aav) as an in vivo gene transfer vector for the retina. | 1997 | 9288458 |
| e4 gene function in adenovirus, adenovirus vector and adeno-associated virus infections. | | 1997 | 9291998 |
| [virally mediated gene transfer in the patellar tendon. an experimental study in rabbits]. | growth factors have the potential to enhance native repair responses in ligamentous and meniscal lesions. however, methods for applying these cytokines to sites of injury for extended periods are lacking. we suggest that local transfer of genes that encode the relevant healing factors merits investigation as a potential solution to this problem. in the present study, different viral vectors and liposomes are evaluated for their ability to deliver genes to cells of ligamentous and meniscal origin ... | 1997 | 9297243 |
| adeno-associated virus rep78 inhibits oncogenic transformation of primary human keratinocytes by a human papillomavirus type 16-ras chimeric. | seroepidemiologic studies demonstrate that adeno-associated virus (aav) infection is negatively associated with cervical cancer. this inverse association may be due to an ability of aav to inhibit the role of human papillomavirus (hpv) in cervical carcinogenesis. in support of this hypothesis aav has been demonstrated to inhibit several papillomavirus types, including bovine papillomavirus type 1 and human papillomaviruses types 16 and 18 (hpv-16/18) in tissue culture. the aav-encoded rep78 prot ... | 1997 | 9299265 |
| the rep68 protein of adeno-associated virus type 2 increases rna levels from the human cytomegalovirus major immediate early promoter. | the rep68 and rep78 proteins of adeno-associated virus type-2 (aav) are multifunctional dna binding proteins which are involved in the positive and negative regulation of aav genes, as well as various cellular and heterologous viral genes. in this study we report that rep68 enhances expression from the major immediate early promoter (miep) of human cytomegalovirus (hcmv). this rep-mediated enhancement of rna levels is abrogated by the introduction of a rep recognition sequence (rrs) at either po ... | 1997 | 9299629 |
| recombinant adeno-associated virus delivers human factor ix in mice. | | 1997 | 9302683 |
| adeno-associated virus type 2 vector for transduction of hematopoietic cells. | | 1998 | 9304711 |
| encapsidation of adeno-associated virus type 2 rep proteins in wild-type and recombinant progeny virions: rep-mediated growth inhibition of primary human cells. | the adeno-associated virus type 2 (aav) arrests the growth of primary human fibroblasts in vitro at high particle-to-cell ratios. to test the role of aav gene expression in the observed growth inhibition, primary human cells were infected, under identical conditions, with wild-type (wt) aav or with recombinant aav that lacked all viral promoters and coding sequences. significant, dose-dependent growth inhibition of primary human cells was observed with both wt and recombinant aav at particle-to- ... | 1997 | 9311814 |
| adeno-associated virus rep proteins target dna sequences to a unique locus in the human genome. | we have developed a system for site-specific dna integration in human cells, mediated by the adeno-associated virus (aav) rep proteins. in its normal lysogenic cycle, aav integrates at a site on human chromosome 19 termed aavs1. we describe a rapid pcr assay for the detection of integration events at aavs1 in whole populations of cells. using this assay, we determined that the aav rep proteins, delivered in cis or trans, are required for integration at aavs1. only the large forms of the rep prot ... | 1997 | 9311886 |
| genetic immunization with adeno-associated virus vectors expressing herpes simplex virus type 2 glycoproteins b and d. | intramuscular injection of mice with an adeno-associated virus (aav) vector expressing herpes simplex virus type 2 glycoprotein b led to the generation of both gb-specific major histocompatibility complex class i-restricted cytotoxic t lymphocytes and anti-gb antibody. aav-mediated immunization was more potent than plasmid dna or protein in generating antibody responses. | 1997 | 9311887 |
| high mobility group chromosomal protein 1 binds to the adeno-associated virus replication protein (rep) and promotes rep-mediated site-specific cleavage of dna, atpase activity and transcriptional repression. | high mobility group protein 1 (hmg1) is an abundant non-histone chromosomal protein which plays a role in several nuclear events involving dna. here we demonstrate that hmg1 physically interacts with the human adeno-associated virus (aav) rep protein. hmg1 promotes the formation of rep-dna complexes and stimulates the activity of rep in site- and strand-specific cleavage of dna and the hydrolysis of atp, functions required for viral gene regulation, replication and site-specific integration of v ... | 1997 | 9312052 |
| adeno-associated virus-mediated gene transfer into rat carotid arteries. | gene transfer into the arterial wall may allow study of the role of specific genes in vascular pathophysiology and development of local gene therapies for vascular disorders. the feasibility of adeno-associated virus (aav)-mediated gene transfer into isolated segments of normal and balloon-injured rat carotid arteries was studied using a recombinant aav carrying cmvlacz as a reporter gene. approximately 10(6) and 10(7) infectious units (iu) of aav were infused into 1 cm isolated segments of the ... | 1997 | 9338002 |
| adeno-associated virus type 2-mediated transduction in primary human bone marrow-derived cd34+ hematopoietic progenitor cells: donor variation and correlation of transgene expression with cellular differentiation. | although the adeno-associated virus type 2 (aav) is known to possess a broad host range that transcends the species barrier, we suggested in an earlier study that aav infection of human cells is receptor mediated (s. ponnazhagan et al., j. gen. virol. 77:1111-1122, 1996). in the present studies, we investigated the ability of aav to infect primary human hematopoietic progenitor cells capable of multilineage differentiation. bone marrow-derived cd34+ cells from 12 hematologically normal volunteer ... | 1997 | 9343178 |
| adeno-associated virus vector integration junctions. | vectors derived from adeno-associated virus (aav) have the potential to stably transduce mammalian cells by integrating into host chromosomes. despite active research on the use of aav vectors for gene therapy, the structure of integrated vector proviruses has not previously been analyzed at the dna sequence level. studies on the integration of wild-type aav have identified a common site-specific integration locus on human chromosome 19; however, most aav vectors do not appear to integrate at th ... | 1997 | 9343199 |
| 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 |
| is gene therapy in cystic fibrosis a realistic expectation? | to date there are 11 human protocols either ongoing or approved for gene therapy for cystic fibrosis (cf) in the united states. there are also two protocols in the united kingdom and one in france. of these, results have been published in four. the protocols vary in the cells targeted, the vectors used, and the frequency of administration, but despite these differences all have contributed toward answering the key questions that will determine the future of gene therapy for cf: the questions of ... | 1996 | 9363187 |
| presence of integrated dna sequences of adeno-associated virus type 2 in four cell lines of human embryonic origin. | the human helper virus-dependent parvovirus adeno-associated virus (aav) has been found in human female genital tissues including material from first trimester miscarriage. in the latter case, aav type 2 (aav-2) dna and viral proteins were detected mainly in the trophoblast cell layer of placenta. in this report, we present evidence that aav dna is also present in established human trophoblast cell lines (jeg-3, jar, bewo) and in the human amnion cell line fl. in cells of these lines, aav-2 dna ... | 1997 | 9367391 |