Publications

ubiquitination of both adeno-associated virus type 2 and 5 capsid proteins affects the transduction efficiency of recombinant vectors.	in the presence of complementing adeno-associated virus type 2 (aav-2) rep proteins, aav-2 genomes can be pseudotyped with the aav-5 capsid to assemble infectious virions. using this pseudotyping strategy, the involvement of the ubiquitin-proteasome system in aav-5 and aav-2 capsid-mediated infections was compared. a recombinant aav-2 (raav-2) proviral luciferase construct was packaged into both aav-2 and aav-5 capsid particles, and transduction efficiencies in a number of cell lines were compar ...	2002	11836382
endocytosis of adeno-associated virus type 5 leads to accumulation of virus particles in the golgi compartment.	among the adeno-associated virus (aav) serotypes which are discussed as vectors for gene therapy aav type 5 (aav5) represents a candidate with unique advantages. to further our knowledge on aav5-specific characteristics, we studied the entry pathway of wild-type virus in hela cells in the absence of helper virus by immunofluorescence and electron microscopy and by western blot analysis. we found virus binding at the apical cell surface, especially at microvilli and, with increasing incubation ti ...	2002	11836412
recent advances in recombinant adeno-associated virus vector production.	adeno-associated virus (aav) is a replication-defective parvovirus that is being developed as a vector for human gene transfer. recombinant aav (raav) vectors are being proposed as a gene transfer vehicle for an array of human diseases. the recent interest in raav has been driven by the unexpected finding that these simple vectors can efficiently transduce a variety of postmitotic cells, resulting in long-lived, robust gene expression. however, a major obstacle to commonplace usage of raav vecto ...	2002	11841606
adeno-associated virus effectively mediates conditional gene modification in the brain.	the cre/loxp system is increasingly showing promise for investigating genes involved in neural function. here, we demonstrate that in vivo modification of genes in the mouse brain can be accomplished in a spatial- and temporal-specific manner by targeted delivery of an adeno-associated virus (aav) encoding a green fluorescent protein/cre recombinase (gfp/cre) fusion protein. by using a reporter mouse, in which cre recombinase activates beta-galactosidase expression, we demonstrate long-term reco ...	2002	11842206
topors, a p53 and topoisomerase i binding protein, interacts with the adeno-associated virus (aav-2) rep78/68 proteins and enhances aav-2 gene expression.	the adeno-associated virus type 2 (aav-2) rep proteins are essential for aav dna replication and regulation of aav gene expression. we have identified a cellular protein interacting with rep78 and rep68 in yeast two-hybrid analysis and in gst pull-down assays. this protein has recently been described as both a p53 (p53bp3) and a topoisomerase i interacting protein (topors). it contains an arginine/serine-rich domain, a ring finger domain and five pest sequences. a minimal sequence sufficient for ...	2002	11842245
inhibition of atherosclerosis in apolipoprotein-e-deficient mice following muscle transduction with adeno-associated virus vectors encoding human apolipoprotein-e.	apolipoprotein e (apoe) is a multifunctional plasma glycoprotein involved in lipoprotein metabolism and a range of cell signalling phenomena. apoe-deficient (apoe(-/-)) mice exhibit severe hypercholesterolaemia and are an excellent model of human atherosclerosis. apoe somatic gene transfer and bone marrow transplantation in apoe(-/-) mice results in reversal of hypercholesterolaemia, inhibition of atherogenesis and regression of atherosclerotic plaque density. replication defective adeno-associa ...	2002	11850719
kinetics of transgene expression in mouse retina following sub-retinal injection of recombinant adeno-associated virus.	using confocal microscopy we have examined in detail the temporal and spatial pattern of green fluorescent protein expression following sub-retinal injection of recombinant adeno-associated virus (raav) in the mouse and have determined the effect of viral titre on the number and type of cells transduced. our results suggest that some transgene expression occurs as early as three days after injection, and that transgene expression occurs beyond the area of retinal detachment. vector titre appears ...	2002	11853771
cftr with a partially deleted r domain corrects the cystic fibrosis chloride transport defect in human airway epithelia in vitro and in mouse nasal mucosa in vivo.	in developing gene therapy for cystic fibrosis (cf) airways disease, a transgene encoding a partially deleted cf transmembrane conductance regulator (cftr) cl- channel could be of value for vectors such as adeno-associated virus that have a limited packaging capacity. earlier studies in heterologous cells indicated that the cftr r (regulatory) domain is predominantly random coil and that parts of the r domain can be deleted without abolishing channel function. therefore, we designed a series of ...	2002	11854474
antitumor effect of an adeno-associated virus vector containing the human interferon-beta gene on experimental intracranial human glioma.	we constructed an adeno-associated virus (aav) vector containing the human interferon-beta (huifn-b ) gene (aav-ifn-beta) and investigated its antitumor effect against human glioma cells (u251-sp) inoculated into the brain of nude mice. prior to this, we examined human glioma cells transduced with aav-ifn-beta using video-enhanced contrast differential interference contrast (vec-dic) microscopy. infection of aav-ifn-beta induced apoptosis and secondary necrosis in human glioma cells. in in vivo ...	2002	11856487
behavioral recovery in a primate model of parkinson's disease by triple transduction of striatal cells with adeno-associated viral vectors expressing dopamine-synthesizing enzymes.	one potential strategy for gene therapy of parkinson's disease (pd) is the local production of dopamine (da) in the striatum induced by restoring da-synthesizing enzymes. in addition to tyrosine hydroxylase (th) and aromatic-l-amino-acid decarboxylase (aadc), gtp cyclohydrolase i (gch) is necessary for efficient da production. using adeno-associated virus (aav) vectors, we previously demonstrated that expression of these three enzymes in the striatum resulted in long-term behavioral recovery in ...	2002	11860702
adeno-associated virus mediated expression of human erythropoietin in vitro.	to investigate the expression of human erythropoietin (hepo) in 293 cell line using adeno-associated virus (aav) vector, and provide information for its potential application in human gene therapy.	2002	11860738
enhanced phagemid particle gene transfer in camptothecin-treated carcinoma cells.	engineered phage-based vectors are an attractive alternative strategy for gene delivery because they possess no natural mammalian cell tropism and can be genetically modified for specific applications. genotoxic treatments that increase the transduction efficiency of single-stranded adeno-associated virus were tested on cells transfected by single-stranded phage. indeed, green fluorescent protein transgene expression by epidermal growth factor-targeted phagemid particles increased with heat shoc ...	2002	11861367
[somatic gene therapy of dilated cardiomyopathy].	the hereditary form of dilated cardiomyopathy (dcm) accounts for about 20% of human dcm and is a major cause of heart failure. to-2 strain hamsters show dcm, a gene deletion of delta-sarcoglycan (sg), loss of all four sgs, alpha-, beta-, gamma- and delta-sg proteins, and are useful for developing gene therapy of the hereditary dcm. the delta-sg is a component of dystrophin-associated glycoprotein complex that stabilizes sarcolemma. four familial and sporadic dcm cases have been reported in human ...	2002	11862755
inducible adeno-associated virus vector-delivered transgene expression in corneal endothelium.	to investigate whether recombinant adeno-associated virus (raav) vector--mediated transgene expression is induced by inflammation in corneal endothelial cells in vivo.	2002	11867594
cross-packaging of a single adeno-associated virus (aav) type 2 vector genome into multiple aav serotypes enables transduction with broad specificity.	the serotypes of adeno-associated virus (aav) have the potential to become important resources for clinical gene therapy. in an effort to compare the role of serotype-specific virion shells on vector transduction, we cloned each of the serotype capsid coding domains into a common vector backbone containing aav type 2 replication genes. this strategy allowed the packaging of aav2 inverted terminal repeat vectors into each serotype-specific virions. each of these helper plasmids (pxr1 through pxr5 ...	2002	11752169
efficient generation of cytotoxic t lymphocytes against cervical cancer cells by adeno-associated virus/human papillomavirus type 16 e7 antigen gene transduction into dendritic cells.	adeno-associated virus (aav) is able to efficiently deliver a cytokine gene into dendritic cells (dc). improvements in t cell priming by dc might be effected by the delivery of antigen genes into dc, resulting in continuous protein expression, as most proteins have short half-lives. in this study, a recombinant aav vector containing the human papillomavirus (hpv)-16 e7 gene was used to pulse/infect dc and compared to the pulsing of dc by the lipofection of bacterially produced e7 protein. pulsin ...	2002	11754001
adeno-associated virus type 2 rep78 inhibition of pka and prkx: fine mapping and analysis of mechanism.	hormones and neurotransmitters utilize cyclic amp (camp) as a second messenger in signal transduction pathways to regulate cell growth and division, differentiation, gene expression, and metabolism. adeno-associated virus type 2 (aav-2) nonstructural protein rep78 inhibits members of the camp signal transduction pathway, the protein kinases pka and prkx. we mapped the kinase binding and inhibition domain of rep78 for prkx to amino acids (aa) 526 to 561 and that for pka to aa 526 to 621. these po ...	2002	11773379
a high-capacity, capsid-modified hybrid adenovirus/adeno-associated virus vector for stable transduction of human hematopoietic cells.	to achieve stable gene transfer into human hematopoietic cells, we constructed a new vector, deltaad5/35.aav. this vector has a chimeric capsid containing adenovirus type 35 fibers, which conferred efficient infection of human hematopoietic cells. the deltaad5/35.aav vector genome is deleted for all viral genes, allowing for infection without virus-associated toxicity. to generate high-capacity deltaad5/35.aav vectors, we employed a new technique based on recombination between two first-generati ...	2002	11773389
adenovirus and adeno-associated virus vectors.	recombinant adenovirus (rad) and recombinant adeno-associated virus (raav) are among the most extensively used vectors in gene therapy studies to date. these two vectors share some similar features such as a broad host range and ability to infect both proliferating and quiescent cells. however, they also possess their own unique set of properties that render them particularly attractive for gene therapy applications. rad vectors can accommodate larger inserts, mediate transient but high levels o ...	2002	12573049
obesity and metabolic syndrome: long-term benefits of central leptin gene therapy.	the recent rapid rise in the incidence of obesity has prompted investigations into understanding the hormonal and neuronal pathways involved in body weight homeostasis in order to devise novel therapeutic strategies. the early enthusiasm for the adipocyte hormone leptin as a regulator of fat mass was largely discarded because of the apparent development of leptin resistance, as seen in obese subjects with elevated blood leptin levels. we postulated that this leptin ineffectiveness may be caused ...	2002	12582458
safety of adeno-associated virus gene therapy vectors: a current evaluation.	an increasing number of strategies for molecular treatment of disease rely on the adeno-associated virus (aav) as a therapeutic gene delivery vector. one of the most attractive features of this viral dna vector is the perceived safety of aav gene delivery. recent applications in human clinical trials support the safety record established in preclinical trials, with evidence of gene transfer in the absence of cellular immune responses or tissue disturbance. nevertheless, many aspects of the biolo ...	2002	12904163
a preliminary evaluation of recombinant adeno-associated virus biodistribution in rhesus monkeys after intrahepatic inoculation in utero.	the ability to deliver genes to fetuses in utero may prove crucial for those genetic diseases that are associated with severe fetal morbidity and for which there is no effective postnatal therapy. in utero therapy may be especially useful in diseases that affect the central nervous system because the immature blood-brain barrier may facilitate gene delivery to neural target cells. we investigated whether in utero inoculation of recombinant adeno-associated virus (raav) into rhesus monkey fetuses ...	2002	12489998
intraarticular gene transfer of tnfr:fc suppresses experimental arthritis with reduced systemic distribution of the gene product.	tumor necrosis factor alpha (tnfalpha) plays a pivotal role in the pathogenesis of rheumatoid arthritis (ra). blockage of tnfalpha actions by systemic administration of tnf antagonists has recently been shown to ameliorate joint symptoms in ra patients. in the present study, a streptococcal cell wall (scw)-induced rat arthritis model was used to evaluate the effect of different gene transfer routes of a tnf antagonist on the development and severity of arthritis. successful delivery of a plasmid ...	2002	12498769
[construction of adeno-associated virus vector carried mutated dihydrofolate reductase and green fluorescent protein and its expression in nih3t3 cells].	the aim of this study was to construct recombinant mdhfr-gfp/aav vector containing mutated dihydrofolate reductase (mdhfr) and green fluorescent protein (gfp) fusion genes and its expression in nih3t3 cells, to investigate the resistance of the cells to methotrexate. amplified cdna of mdhfr and gfp segmented from their plasmid separately were linked by pcr with the aminoacetic acid linker. the fusion gene was inserted into t vector, and after enzyme cutting the fusion gene fragment was inserted ...	2002	12513788
[effect of antisense thrombin receptor and p21 double gene co-expression system on the proliferation and apoptosis in human aortic smooth muscle cells].	to focus on the study of the effect on proliferation and apoptosis of human aortic smooth muscle cells (asmc) by adeno-associated virus (aav) vector carrying antisense thrombin receptor (atr) and p21 double gene co-expression system.	2002	12905648
retroviral-mediated transduction and clonal integration analysis of human hematopoietic stem and progenitor cells.	this chapter provides information on the methods used to introduce genes into human hematopoietic stem and progenitor cells, using moloney murine leukemia (momulv)-based retroviral vectors. momulv-based vectors have the ability to efficiently transfer genes into mammalian cells, leading to permanent integration of a single copy of the gene of interest into the cellular chromosomes. the technique of single-colony inverse [polymerase chain reaction (pcr) can be used to track individual descendants ...	2002	21437813
a novel and highly efficient production system for recombinant adeno-associated virus vector.	recombinant adeno-associated virus (raav) has proven to be a promising gene delivery vector for human gene therapy. however, its application has been limited by difficulty in obtaining enough quantities of high-titer vector stocks. in this paper, a novel and highly efficient production system for raav is described. a recombinant herpes simplex virus type 1 (rhsv-1) designated hsv1-rc/deltaul2, which expressed adeno-associated virus type2 (aav-2) rep and cap proteins, was constructed previously. ...	2002	18763068
sarcolemmal fragility secondary to the degradation of dystrophin in dilated cardiomyopathy, as estimated by electron microscopy.	a common gene deletion or mutation of delta-sarcoglycan (delta-sg) in dystrophin-related proteins (drps) is identified in both to-2 strain hamsters and human families with dilated cardiomyopathy. we have succeeded in the long-lasting in vivo supplementation of a normal delta-sg gene by recombinant adeno-associated virus vector, restoration of the morphological and functional degeneration, and improvement in the prognosis of the to-2 hamster. to evaluate the integrity of the sarcolemma (sl) and t ...	2003	19641652
prospects for the treatment of stroke using gene therapy.	recent advances have demonstrated the use of gene therapy in the treatment of stroke in experimental animal models of focal ischemia, global ischemia and subarachnoid hemorrhage. several different vectors for gene transfer have been studied including herpes simplex virus, adenovirus, adeno-associated virus and liposomes. genetically modified cell lines (e.g., bone marrow-derived cells) have been studied for ex vivo gene therapy. the effects of gene transfer to several brain regions including the ...	2003	19810903
nanoparticle-directed tissue-specific delivery system for genes and drugs.	extract: gene therapy is recognized as a promising approach for the treatment of serious diseases including monogenic diseases, infectious diseases, and cancer. conventional vectors used for delivering therapeutic genes are virus-based vectors, such as the adenovirus, retrovirus, lentivirus, and adeno-associated virus. since these viruses can infect a wide range of tissues, it is essential to be able to deliver these viruses specifically to the target tissues in vivo. if these viruses are admini ...	2003	20704869
modified adeno-associated virus (aav) vectors: a new generation of targeted gene therapy delivery systems.	extract: gene therapy in its simplest form is the deliberate transfer of therapeutic genes into a host cell. as a human gene therapy vector, adeno-associated virus (aav) is a promising delivery system. it is able to maintain stable gene expression in host cells, is efficient at gene delivery in vivo, and is non-pathogenic. however, one obstacle facing the advancement of aav vectors is the natural tropism of the virus. the primary attachment receptor for aav type-2 (aav2) is heparan sulfate prote ...	2003	20705032
non-small lung cancer cells are prime targets for p53 gene transfer mediated by a recombinant adeno-associated virus type-2 vector.	in this study, we elucidated the potential of recombinant adeno-associated virus type-2 (raav-2) vectors for lung cancer gene therapy. cell lines of the three major histological subtypes of non-small cell lung cancer (nsclc) were highly susceptible for raav-2 showing transduction rates between 63.4 and 98.9%. in contrast, cell lines of small cell carcinomas were resistant to raav-2 infection. for restoration of p53 function in p53 deficient nsclc, a raav-2 vector was constructed containing wt p5 ...	2003	14712316
[generation of human recombinant antibody fab fragment and its igg to adeno-associated virus type ii from phage display library].	to acquire the recombinant human monoclonal antibodies and igg to adeno-associated virus type 2 (aavs-2).	2003	15340567
intramuscular injection of aav-gdnf results in sustained expression of transgenic gdnf, and its delivery to spinal motoneurons by retrograde transport.	adeno-associated virus (aav) vector has been developed as an attractive gene delivery system with proven safety. glial cell line-derived neurotrophic factor (gdnf) is proposed to be a promising therapeutic agent for amyotrophic lateral sclerosis (als) and other motor neuron diseases. the purpose of this report was to investigate transgenic gdnf expression at different time points post aav mediated gdnf intramuscular delivery. an aav vector was constructed to encode a recombinant fusion of gdnf t ...	2003	12507722
[inhibition of er alpha-mannosidase expression causes reduction and shortening of microvilli on rat liver epithelial cell wb-f344].	to study the effect of n-glycosylation on the modification of microvilli on the surface of rat liver epithelial cell wb-f344 and the growth of the cells in culture.	2003	12905608
a novel recombinant adeno-associated virus vaccine reduces behavioral impairment and beta-amyloid plaques in a mouse model of alzheimer's disease.	memory impairment progressing to dementia is the main clinical symptom of alzheimer's disease (ad). deposition of the amyloid-beta peptide (abeta) in brain, particularly its 42-amino acid isoform (abeta42), has been shown to play a primary and crucial role in the pathogenesis of ad. in this study we have developed a recombinant adeno-associated virus (aav) vaccine against ad. this vaccine could express cb-abeta42 (cholera toxin b subunit and abeta42 fusion protein) in vivo. a single administrati ...	2003	14678754
glucose-responsive expression of the human insulin promoter in hepg2 human hepatoma cells.	the concept of insulin production afforded by hepatic gene therapy retains promise as a potential therapy for type 1 diabetes, but the approach has been limited by the need for strict transgene regulation in response to fluctuating levels of both glucose and insulin. furthermore, while hepatocytes contain various glucose-responsive elements, they lack the appropriate regulated secretory system necessary for insulin release, thereby necessitating the requirement for transcriptional regulation of ...	2003	14679068
evaluation of risks related to the use of adeno-associated virus-based vectors.	recombinant aav efficacy has been demonstrated in numerous gene therapy preclinical studies. as this vector is increasingly applied to human clinical trials, it is a priority to evaluate the risks of its use for workers involved in research and clinical trials as well as for the patients and their descendants. at high multiplicity of infection, wild-type aav integrates into human chromosome 19 in approximately 60% of latently infected cell lines. however, it has been recently demonstrated that o ...	2003	14683451
[transduction efficiency of recombinant adeno-associated virus 2 in human umbilical cord blood cd34+ hematopoietic stem/progenitor cells].	to investigate the transduction efficiency of recombinant adeno-associated virus 2 (raav-2) in human umbilical cord blood cd34(+) hematopoietic stem/progenitor cells, the cd34(+) cells sorted by the method of magnetic cell sorting from human cord blood were infected with the raav-2 expressing the green fluorescent protein (gfp) gene. after transduction for 19 hours, the expression of gfp was detected under fluorescence microscope. the results showed that 43% cd34(+) cells expressed the gfp gene ...	2003	14706137
the suppressor of cytokine signaling-1 (socs1) is a novel therapeutic target for enterovirus-induced cardiac injury.	enteroviral infections of the heart are among the most commonly identified causes of acute myocarditis in children and adults and have been implicated in dilated cardiomyopathy. although there is considerable information regarding the cellular immune response in myocarditis, little is known about innate signaling mechanisms within the infected cardiac myocyte that contribute to the host defense against viral infection. here we show the essential role of janus kinase (jak) signaling in cardiac my ...	2003	12588885
long-term liver allograft survival induced by combined treatment with raav-hctla4ig gene transfer and low-dose fk506.	recombinant adeno-associated virus vector (raav) is a promising vehicle for gene delivery, but few reports have documented its application in solid organ transplantation. in a rat orthotopic liver transplantation model, we investigated the efficacy of raav-mediated human cytotoxic t-lymphocyte-associated antigen 4 and immunoglobulin g (hctla4ig) gene transfer to induce long-term allograft survival.	2003	12589149
treatment of experimental asthma by long-term gene therapy directed against il-4 and il-13.	the clinical manifestations of allergic asthma are believed to result from a dysregulated, t helper 2 lymphocyte (th2)-biased response to antigen. although asthma symptoms can be controlled acutely, there is a need for a therapy that will address the underlying immune dysfunction and provide continuous control of chronic airway inflammation. the th2-type cytokines, il-13 and il-4, have been demonstrated to play a crucial role in asthma pathogenesis and their selective neutralization results in t ...	2003	12597903
long-term correction of glycogen storage disease type ii with a hybrid ad-aav vector.	we administered an adenovirus-adeno-associated virus (ad-aav) vector encoding human acid alpha-glucosidase (hgaa) to acid alpha-glucosidase-knockout (gaa-ko) mice on day 3 of life by gastrocnemius injection. in contrast to previous results for muscle-targeted ad vector in adult gaa-ko mice, the muscles of the hindlimb showed reduced glycogen content and persistent hgaa for as long as 6 months after neonatal ad-aav vector administration. not only the injected gastrocnemius muscles, but also the h ...	2003	12597907
optic neuropathy induced by reductions in mitochondrial superoxide dismutase.	reactive oxygen species (ros) are suspected to play a pivotal role in the pathogenesis of leber hereditary optic neuropathy (lhon), caused by mutated complex i subunit genes. it seems surprising that optic neuropathy has not been described in animals with a knockout of genes encoding critical anti-ros defenses. if ros have a role in the optic nerve injury of lhon, then increasing mitochondrial levels of ros should induce optic neuropathy.	2003	12601034
nigrostriatal alpha-synucleinopathy induced by viral vector-mediated overexpression of human alpha-synuclein: a new primate model of parkinson's disease.	we used a high-titer recombinant adeno-associated virus (raav) vector to express wt or mutant human alpha-synuclein in the substantia nigra of adult marmosets. the alpha-synuclein protein was expressed in 90-95% of all nigral dopamine neurons and distributed by anterograde transport throughout their axonal and dendritic projections. the transduced neurons developed severe neuronal pathology, including alpha-synuclein-positive cytoplasmic inclusions and granular deposits; swollen, dystrophic, and ...	2003	12601150
efficient trans-splicing in the retina expands the utility of adeno-associated virus as a vector for gene therapy.	recombinant vectors based on adeno-associated virus (aav) can efficiently transduce many different cell types, including cells of the retina, resulting in stable gene expression. a major shortcoming of this vector is its small packaging capacity. a trans-splicing approach, which reconstitutes gene expression from two independent aav vectors, can be used to overcome the vector's packaging limitations. the efficiency of this system to date has been disappointing, and therefore its utility for ther ...	2003	12573057
helper-independent and aav-itr-independent chromosomal integration of double-stranded linear dna vectors in mice.	nonviral plasmid dna is a promising vector for achieving ex vivo and in vivo gene transfer. however, transgene expression is usually transient, especially in dividing target cells due to loss of vector genomes. here we describe the use of naked double-stranded (ds) linear dna as a way to insert exogenous dna sequences into chromosomes of mouse hepatocytes in vivo, without helper components such as integrase or transposase. we constructed ds linear dna vectors with or without adeno-associated vir ...	2003	12573623
free dna ends are essential for concatemerization of synthetic double-stranded adeno-associated virus vector genomes transfected into mouse hepatocytes in vivo.	recombinant adeno-associated virus (raav) vectors stably transduce hepatocytes in vivo. in hepatocyte nuclei, the incoming single-stranded (ss) vector genomes are converted into various forms of double-stranded (ds) genomes including extrachromosomal linear and circular monomers and concatemers, and a small portion of the vector genomes integrate into chromosomes. the mechanism of genome conversion is not well understood. in the present study, we analyzed the role of inverted terminal repeat (it ...	2003	12573624
quantification of adeno-associated virus particles and empty capsids by optical density measurement.	we show here that uv absorbance of denatured adeno-associated virus (aav) vector provides a simple, rapid, and direct method for quantifying vector genomes and capsid proteins in solution. we determined the molar extinction coefficients of capsid protein to be 3.72 x 10(6) m(-1) cm(-1) at 260 nm and 6.61 x 10(6) m(-1) cm(-1) at 280 nm. for recombinant aav vectors, extinction coefficients can be calculated by including the predicted absorbance of the vector dna. since the amount of empty capsids ...	2003	12573625
enhancement of gene transfer with recombinant adeno-associated virus (raav) vectors into primary b-cell chronic lymphocytic leukemia cells by cpg-oligodeoxynucleotides.	transduction of primary b-cell chronic lymphocytic leukemia (b-cll) cells with recombinant adeno-associated virus (raav) vectors is dependent on preactivation of leukemic cells by cd40l. cpg-oligodeoxynucleotides (cpg-odns) are able to activate cytokine production and proliferation of b-cll cells. therefore cpg-odns were tested for their potential to enhance transgene expression in cll cells.	2003	14662328
persistent phenotypic correction of central diabetes insipidus using adeno-associated virus vector expressing arginine-vasopressin in brattleboro rats.	adeno-associated virus (aav) vector is suitable for gene transfer to the central nervous system. however, the efficacy of gene therapy for neuroendocrine disease is still unknown. in this study, we injected aav vector encoding arginine-vasopressin (avp) stereotaxically into the bilateral hypothalamus of brattleboro rats. brattleboro rats show a central diabetes insipidus (cdi) phenotype and growth retardation due to a complete deficiency of avp. following injection, both urine volume and urine o ...	2003	14664791
adeno-associated virus-mediated gene transfer of a secreted decoy human macrophage scavenger receptor reduces atherosclerotic lesion formation in ldl receptor knockout mice.	macrophage scavenger receptors (msr) promote atherosclerotic lesion formation, and modulation of msr activity has been shown to influence atherosclerosis. soluble receptors are effective in inhibiting receptor-mediated functions in various diseases. we have generated a secreted macrophage scavenger receptor (smsr) that consists of the bovine growth hormone signal sequence and the human msr a i extracellular domains. smsr reduces degradation of atherogenic modified low-density lipoproteins and mo ...	2003	14664792
self-complementary adeno-associated virus serotype 2 vector: global distribution and broad dispersion of aav-mediated transgene expression in mouse brain.	the blood-brain barrier is the main obstacle to efficient delivery of therapeutic reagents, including viral vectors, into the central nervous system (cns) for treating global cns diseases. in this study, the effects of mannitol infusions on global brain gene expression of a novel aav vector were examined after intravenous (i.v.) or intracisternal injection. initially, a self-complementary adeno-associated virus serotype 2 vector (scaav) was compared to traditional single-stranded aav2 vector for ...	2003	14664793
fibroblast growth factor-2 gene delivery stimulates axon growth by adult retinal ganglion cells after acute optic nerve injury.	basic fibroblast growth factor (or fgf-2) has been shown to be a potent stimulator of retinal ganglion cell (rgc) axonal growth during development. here we investigated if fgf-2 upregulation in adult rgcs promoted axon regrowth in vivo after acute optic nerve injury. recombinant adeno-associated virus (aav) was used to deliver the fgf-2 gene to adult rgcs providing a sustained source of this neurotrophic factor. fgf-2 gene transfer led to a 10-fold increase in the number of axons that extended p ...	2003	14664816
comparison of murine leukemia virus, human immunodeficiency virus, and adeno-associated virus vectors for gene transfer in multiple myeloma: lentiviral vectors demonstrate a striking capacity to transduce low-proliferating primary tumor cells.	genetic modification of primary tumor cells by gene transfer is of major interest to study the role of specific genes in the biology of a given malignancy and to modify tumor cells for therapeutic use. multiple myeloma (mm) is a low-proliferating cancer, with often less than 1% of the cells in the s phase of the cell cycle. as primary myeloma cells are notoriously difficult to transduce, we conducted a comparison of various viral vectors, known to integrate the transgene of interest into the tar ...	2003	14670124
ectopic expression of the trka receptor in adult dopaminergic mesencephalic neurons promotes retrograde axonal ngf transport and ngf-dependent neuroprotection.	a recombinant adeno-associated virus (raav) was used to investigate the impact of an ectopic expression of the ngf high-affinity receptor in adult neurons. the rat trka cdna cloned in a pcmx vector was first tagged with a human c-myc sequence. the resulting vector was shown to encode a functional receptor which promoted the expression of trka immunoreactivity upon transfection of 293 fibroblasts or nnr5 cells, a trka-defective variant of pc12 cells. these cells also accumulate trka transcripts u ...	2003	14552878
rescue of the adeno-associated virus genome from a plasmid vector: evidence for rescue by replication.	in cultured cells, adeno-associated virus (aav) replication requires coinfection with a helper virus, either adenovirus or herpesvirus. in the absence of helper virus coinfection aav can integrate its genome site specifically into the aavs1 region of chromosome 19. upon subsequent infection with a helper virus, the aav genome is released from chromosome 19 by a process termed rescue, and productive replication ensues. the aav genome cloned into a plasmid vector can also serve to initiate product ...	2003	14557633
prolonged survival of heart allografts transduced with aav-ctla4ig.	organ grafts transduced with gene-encoding immunosuppressive molecules are a less toxic approach to preventing graft rejection. adenovirus vectors have been widely tested with unsatisfactory results, while adeno-associated virus (aav) is smaller and elicits a low host humoral response. we constructed an aav vector containing the mouse ctla4ig gene. b10 (h2(b)) cardiac grafts were transduced with aav-ctla4ig by coronary infusion. aav-lacz vectors were used as reporters and controls, and the expre ...	2003	14558008
identification of new mhc-restriction elements for presentation of the p210(bcr-abl) fusion region to human cytotoxic t lymphocytes.	chronic myelogenous leukemia (cml) is characterized by a t(9;22) translocation resulting in expression of bcr-abl fusion oncoproteins which are unique to the leukemic cells, necessary for oncogenesis, and potentially immunogenic. we have previously shown that human dendritic cells transduced with an adeno-associated virus vector encoding the fusion region of the b3a2 splice variant (p210(b3a2)) of the bcr-abl oncoprotein elicit specific t-cell responses in vitro. two cytotoxic t lymphocyte (ctl) ...	2003	14564482
ssdna-dependent colocalization of adeno-associated virus rep and herpes simplex virus icp8 in nuclear replication domains.	the subnuclear distribution of replication complex proteins is being recognized as an important factor for the control of dna replication. herpes simplex virus (hsv) single-strand (ss)dna-binding protein, icp8 (infected cell protein 8) accumulates in nuclear replication domains. icp8 also serves as helper function for the replication of adeno-associated virus (aav). using quantitative 3d colocalization analysis we show that upon coinfection of aav and hsv the aav replication protein rep and icp8 ...	2003	14576307
long-term neuronal effects and disposition of ectopic preprongf gene transfer into the rat septum.	although ngf gene therapy has been proposed to treat age- or disease-related brain cholinergic decline, little is known about the ectopic expression or function of this trophic factor after transduction in the brain especially over long intervals. the neuron-targeting, recombinant adeno-associated virus serotype 2 (raav2) vector was used to express mouse ngf with c-terminal myc-tag in septum using a full-length preprongf sequence. while the predominant form of endogenous ngf immunoreactivity in ...	2003	14577926
use of localised gene transfer to develop new treatment strategies for the salivary component of sjögren's syndrome.	effective treatment for sjögren's syndrome (ss) might be developed locally by introducing genes encoding cytokines, which are potentially anti-inflammatory, or by introducing a cdna encoding a soluble form of a key cytokine receptor, which can act as an antagonist and decrease the availability of certain cytokines, such as soluble tumour necrosis factor alpha receptors. currently, the preferred choice of viral vector for immunomodulatory gene transfer is recombinant adeno-associated virus. the u ...	2003	14583564
construction of adeno-associated virus packaging plasmids and cells that directly select for aav helper functions.	recombinant adeno-associated virus type 2 (raav) has promise for use as a gene therapy vector. potential problems in the production of raav stocks are both the limited amount of recombinant virus that is produced by traditional methods and the possibility of wild-type replication competent adeno-associated virus (wtaav) contamination. the presence of these contaminants is largely dependent upon the helper plasmid used. whilst wtaav is not a pathogen, the presence of these contaminants is undesir ...	2003	14599673
long-term transgene expression in cardiac and skeletal muscle following fetal administration of adenoviral or adeno-associated viral vectors in mice.	in utero gene transfer may provide advantages for the correction of congenital genetic disorders. in the present study we compare the ability of adenovirus (adcmvlacz), and two serotypes of adeno-associated virus (aavcmvlacz serotypes 2 and 2/5), to target cardiac and skeletal muscle after prenatal systemic or intramuscular injection in mice and assess the immune response to the vectors.	2003	14601131
aav-mediated gene therapy for hemophilia.	gene therapy for hemophilia has been contemplated since the coagulation factor genes responsible for the disease were cloned 20 years ago. multiple approaches towards the delivery of factors viii or ix, the defective genes in the most common forms of hemophilia, have resulted in positive results in animals, and largely equivocal results in human clinical testing. use of vectors based on adeno-associated virus has led to robust and sustained cures in hemophilic mice and dogs, and intriguing preli ...	2003	14601521
intracavernosal vascular endothelial growth factor (vegf) injection and adeno-associated virus-mediated vegf gene therapy and reverse venogenic erectile dysfunction in rats.		2003	14601576
transductional and transcriptional targeting of cancer cells using genetically engineered viral vectors.	gene delivery vectors, including adenovirus (ad) and adeno-associated virus (aav), are inefficient and non-selective for cancer due to low levels of viral receptors with high levels on other tissues, including liver. we tested ads and aavs with the sigyplp-targeting peptide inserted into virus capsids for transduction in a panel of cancer cells. six of twelve lines (c8161, pc-3, g-ccm, mkn-45, lncap and a549) were transduced, independently of native viral tropism. furthermore the candidate cance ...	2003	14607330
predicting the future of human gene therapy for cardiovascular diseases: what will the management of coronary artery disease be like in 2005 and 2010?	gene therapy is the use of gene delivery as a means to achieve high levels of the therapeutic gene product (ie, "drug" delivery) to treat acquired cardiovascular diseases. human gene therapy for cardiovascular disease is expected to provide important advances in therapeutic angiogenesis, myocardial protection, myocardial regeneration and repair, restenosis, prevention of bypass graft failure, and risk-factor management. the data from ongoing phase 2 and future phase 3 studies will provide eviden ...	2003	14615024
sustained peripheral expression of transgene adiponectin offsets the development of diet-induced obesity in rats.	adiponectin (acrp30) is a physiologically active polypeptide hormone secreted by adipose tissue that shows insulin-sensitizing, antiinflammatory, and antiatherogenic properties. in humans, acrp30 levels are inversely related to the degree of adiposity. in the current study, we tested the long-term weight-reducing and insulin-enhancing effects of acrp30 cdna delivered peripherally by a viral vector. to this end, we have generated a series of recombinant adeno-associated virus vectors of serotypes ...	2003	14617771
serum-free production and column purification of adeno-associated virus type 5.	viral vectors derived from adeno-associated virus (aav) are rapidly becoming the vehicles of choice for gene therapy applications. aav-2 is the adeno-associated virus serotype most commonly employed in aav-mediated gene therapy studies; however, recently developed vectors derived from alternative serotypes of aav, such as aav-5, are receiving special attention due to their disparate tissue tropisms and potential for serial administration. in this report, we describe a rapid and efficient method ...	2003	14625046
rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo.	adeno-associated virus (aav) is a promising gene vector based on a single-stranded (ss) dna virus. its transgene expression requires the conversion of ssdna to double-stranded (ds) genome, a slow process responsible for the delayed transduction and occasional inefficiency. by mutating the inverted terminal repeat, we have made novel aav vectors that predominantly package the self-complementary dsdna genome. the dsaav consistently demonstrated superior and accelerated transduction in vitro and in ...	2003	14625564
adeno-associated virus terminal repeat (tr) mutant generates self-complementary vectors to overcome the rate-limiting step to transduction in vivo.	an important limitation of recombinant adeno-associated virus (raav) vector efficiency is the requirement of hostcell-mediated synthesis of double-stranded dna from the single-stranded genome. we have bypassed this step in a specialized self-complementary raav (scaav) vector, by utilizing the tendency of aav to package dna dimers when the replicating genome is half the length of the wild type (wt). to produce these vectors efficiently, we have deleted the terminal resolution site (trs) from one ...	2003	14625565
genetic modifications of the adeno-associated virus type 2 capsid reduce the affinity and the neutralizing effects of human serum antibodies.	the high prevalence of human serum antibodies against adeno-associated virus type 2 (aav) vectors represents a potential limitation for in vivo applications. consequently, the development of aav vectors able to escape antibody binding and neutralization is of importance. to identify capsid domains which contain major immunogenic epitopes, six aav capsid mutants carrying peptide insertions in surface exposed loop regions (i-261, i-381, i-447, i-534, i-573, i-587) were analyzed. two of these mutan ...	2003	14625569
alpha-1-antitrypsin expression in the lung is increased by airway delivery of gene-transfected macrophages.	inadequate antiprotease activity in the lungs due to alpha-1-antitrypsin (a1at) deficiency is a factor of early-onset emphysema. we propose a new approach to gene therapy that involves the intratracheal delivery of macrophages expressing human a1at (ha1at). recombinant adeno-associated virus (raav) plasmids encoding the ha1at gene were packaged into virions using 293 cells, and transgenic progeny virus was purified from the cells. the murine macrophage cell line j774a.1 was infected in vitro wit ...	2003	14625570
local adeno-associated virus-mediated interleukin 10 gene transfer has disease-modifying effects in a murine model of sjögren's syndrome.	female nonobese diabetic (nod) mice develop spontaneous autoimmune sialadenitis and loss of salivary flow, and are a widely used model of sjögren's syndrome. we examined the feasibility of local salivary gland immunomodulatory gene delivery to alter these sequelae in nod mice. we constructed recombinant adeno-associated virus (raav) vectors encoding either human interleukin 10 (raavhil-10) or beta-galactosidase (raavlacz, control vector). mice received raavhil-10 or raavlacz by retrograde subman ...	2003	14633403
adeno-associated virus type 5: transduction efficiency and cell-type specificity in the primate retina.	gene transfer using adeno-associated viruses (aavs) has been effective for treating inherited retinal diseases in animal models. further evaluation in primates must be performed prior to clinical application, however, because of the difference between the retina of the primate and those of other animals. prior work has shown that aav2 can transduce rod-photoreceptor and rpe cells in the non-human primate retina and that aav5 is more efficient at transducing photoreceptor cells than aav2 in the r ...	2003	14633408
robust adenoviral and adeno-associated viral gene transfer to the in vivo murine heart: application to study of phospholamban physiology.	viral gene transfer to the whole heart in vivo has been achieved in several mammalian species but remained difficult to accomplish in murine hearts. we postulated that a key impediment derives from the use of proximal aortic occlusion during virus injection, because this eliminates coronary perfusion gradients in mice as aortic root and left ventricle pressures equalize.	2003	14638552
prospects and risc score of viral gene therapy for sarcoma.	soft tissue sarcomas are a challenge for medical oncology and gene therapy. protective and sensitising approaches that target normal and malignant tissue, respectively, both have their role for opening the therapeutic window. recent data show that an intensive maintenance chemotherapy significantly reduces metastatic spread and improves disease-free survival in selected patient groups. however, delays of treatment due to cytopenia are frequent. cytostatic drug resistance gene transfer to haemato ...	2003	14640950
[reversion of ovarian carcinoma metastasis by adeno-associated virus-mediated gene transfer of nm23h1 in orthotopic implantation model].	to explore the feasibility of reverse effect of recombinant nm23h(1) adeno-associated virus (raav-nm23h(1)) in ovarian carcinoma metastatic orthotopic implantation nude model.	2003	14642101
chromosomal integration and homologous gene targeting by replication-incompetent vectors based on the autonomous parvovirus minute virus of mice.	the molecular mechanisms responsible for random integration and gene targeting by recombinant adeno-associated virus (aav) vectors are largely unknown, and whether vectors derived from autonomous parvoviruses transduce cells by similar pathways has not been investigated. in this report, we constructed vectors based on the autonomous parvovirus minute virus of mice (mvm) that were designed to introduce a neomycin resistance expression cassette (neo) into the x-linked human hypoxanthine phosphorib ...	2003	14645570
crystal structure of the sf3 helicase from adeno-associated virus type 2.	we report here the crystal structure of an sf3 dna helicase, rep40, from adeno-associated virus 2 (aav2). we show that aav2 rep40 is structurally more similar to the aaa(+) class of cellular proteins than to dna helicases from other superfamilies. the structure delineates the expected walker a and b motifs, but also reveals an unexpected "arginine finger" that directly implies the requirement of rep40 oligomerization for atp hydrolysis and helicase activity. further, the rep40 aaa(+) domain is n ...	2003	12906833
aav-mediated gene transfer slows photoreceptor loss in the rcs rat model of retinitis pigmentosa.	in the royal college of surgeons (rcs) rat, the retinal pigment epithelium (rpe) cannot phagocytose the outer segment discs that are continually shed from photoreceptors. the resulting accumulation of debris in the subretinal space leads to a progressive loss of photoreceptors. the defect results from a mutation in the mertk gene, which is normally expressed in the rpe. mertk is a receptor tyrosine kinase, involved in the binding of photoreceptor debris. mutations in mertk have also been describ ...	2003	12907141
retrograde viral delivery of igf-1 prolongs survival in a mouse als model.	amyotrophic lateral sclerosis (als) is a progressive, lethal neuromuscular disease that is associated with the degeneration of spinal and brainstem motor neurons, leading to atrophy of limb, axial, and respiratory muscles. the cause of als is unknown, and there is no effective therapy. neurotrophic factors are candidates for therapeutic evaluation in als. although chronic delivery of molecules to the central nervous system has proven difficult, we recently discovered that adeno-associated virus ...	2003	12907804
targeting recombinant adeno-associated virus vectors to enhance gene transfer to pancreatic islets and liver.	human pancreatic islet cells and hepatocytes represent the two most likely target cells for genetic therapy of type i diabetes. however, limits to the efficiency of raav serotype 2 (raav2)-mediated gene transfer have been reported for both of these cell targets. here we report that nonserotype 2 aav capsids can mediate more efficient transduction of islet cells, with aav1 being the most efficient serotype in murine islets, suggesting that receptor abundance could be limiting. in order to test th ...	2003	12907946
evidence that increased 12-lipoxygenase expression impairs pancreatic beta cell function and viability.	leukocyte type 12-lipoxygenase (12-lo) is an enzyme specifically expressed in the beta cells of the pancreas. 12-lo oxidizes fatty acids such as arachidonic acid and linoleic acids to their respective hydroperoxides. increased concentration of lipid hydroperoxides causes oxidative stress and this could lead to cellular dysfunction. increased expression of 12-lo in beta cells has been observed with use of inflammatory cytokines and during the prediabetic phase of beta cell dysfunction in the zuck ...	2003	12914766
adeno-associated virus-mediated bone morphogenetic protein-4 gene therapy for in vivo bone formation.	adeno-associated virus (aav) is so far the most valuable vehicle for gene therapy because it has no association with immune response and human disease. the present study was conducted to investigate the feasibility of aav-mediated bmp4 gene transfer for bone formation. in vitro study suggested that aav-bmp4 vectors could transduce myoblast c2c12 cells and produce osteogenic bmp4. in vivo study demonstrated that new bone formation could be induced by direct injection of aav-bmp4 into the skeletal ...	2003	12914798
gene transfer of ikappabalpha limits infarct size in a mouse model of myocardial ischemia-reperfusion injury.	nuclear factor-kappab (nf-kappab) plays a central role in myocardial ischemia-reperfusion (mi/r) injury. the inhibitory protein ikappabalpha prevents its activation. we investigated the effects of adeno-associated viral vector-mediated ikappabalpha gene transfer in mi/r injury. male c57bl/6 mice were randomized to receive a recombinant adeno-associated virus (raav) encoding the gene for the nf-kappab inhibitory protein ikappabalpha (raav- ikappabalpha) or the beta-galactosidase gene (a control a ...	2003	12920239
targeted transgene integration into transgenic mouse fibroblasts carrying the full-length human aavs1 locus mediated by hsv/aav rep(+) hybrid amplicon vector.	herpes simplex virus type 1/adeno-associated virus (hsv/aav) rep(+) hybrid amplicon vectors containing aav inverted terminal repeats (itrs) and rep gene sequences can mediate site-specific integration into the human genome. in this study, we have generated and characterized the first transgenic mice that bear the full-length (8.2 kb) human aavs1 locus. immortalized mouse embryonic fibroblasts from this mouse line were transduced with the rep(+), rep(-) (containing only itrs flanking the transgen ...	2003	12923568
systemic overexpression of il-10 induces cd4+cd25+ cell populations in vivo and ameliorates type 1 diabetes in nonobese diabetic mice in a dose-dependent fashion.	early systemic treatment of nonobese diabetic mice with high doses of recombinant adeno-associated virus (raav) vector expressing murine il-10 prevents type 1 diabetes. to determine the therapeutic parameters and immunological mechanisms underlying this observation, female nonobese diabetic mice at 4, 8, and 12 wk of age were given a single i.m. injection of raav-murine il-10 (10(4), 10(6), 10(8), and 10(9) infectious units (iu)), raav-vector expressing truncated murine il-10 fragment (10(9) iu) ...	2003	12928371
[production and purification of helper free recombinant adeno-associated virus 2].	to explore an efficient and simple way to product and purify helper free adeno-associated virus vectors.	2003	12930642
anti-apoptotic effects of cntf gene transfer on photoreceptor degeneration in experimental antibody-induced retinopathy.	autoantibodies against recoverin are found in the sera of patients with cancer-associated retinopathy syndrome, a paraneoplastic disease associated with retinal degeneration. we have previously shown that anti-recoverin autoantibodies induced photoreceptor apoptotic cell death after injection into the vitreous of lewis rats. ciliary neurotrophic factor (cntf) has been shown to promote the survival of a number of neuronal cell types, including photoreceptors. in this study, we examined whether an ...	2003	12935781
long-term restitution of 4-aminopyridine-sensitive currents in kv1dn ventricular myocytes using adeno-associated virus-mediated delivery of kv1.5.	overexpression of a dominant-negative truncated kv1.1 (kv1dn) polypeptide in the mouse heart resulted in marked attenuation of a 4-aminopyridine (4-ap)-sensitive current, i(k,slow1). we used recombinant adeno-associated virus (raav) as a vector for direct delivery of kv1.5 into the mouse myocardium in order to normalize the action potential duration (apd) 6 months after injection. the injection of raav-kv1.5 reconstituted the 4-ap-sensitive outward potassium currents, shortened the apd, and elim ...	2003	12935889
a sensitive, real-time, rna-specific pcr method for the detection of recombinant aav-cftr vector expression.	following adeno-associated virus (aav)-mediated transduction, cellular rna preparations can be contaminated with aav single-stranded dna. the single-stranded dna genome of recombinant aav vectors can serve as an efficient, but undesirable, template for traditional reverse transcriptase-polymerase chain reaction (rt-pcr) methods. consequently, recombinant aav gene therapy presents a unique challenge to the design of sensitive and reliable methods to detect vector-derived mrna. several methods hav ...	2003	12939641
parvovirus host range, cell tropism and evolution.	the past few years have seen major advances in our understanding of the controls of evolution, host range and cell tropism of parvoviruses. notable findings have included the identification of the transferrin receptor tfr as the cell surface receptor for canine parvovirus and feline panleukopenia virus, and also the finding that specific binding to the canine tfr led to the emergence of canine parvovirus as a new pathogen in dogs. the structures of the adeno-associated virus-2 and porcine parvov ...	2003	12941411
differentiation of marrow stromal cells into photoreceptors in the rat eye.	retinal degenerations and dystrophies are the major causes of genetically inherited blindness that are characterized by the apoptotic death of the photoreceptor cell layer of the retina. to date, no treatment exists for these diseases and only recently have they been considered as candidates for gene and stem cell therapies. here we report the ability of adult cd90+ marrow stromal cells (mscs) to be induced by activin a, taurine, and egf into cells (20-32%) expressing photoreceptor-specific mark ...	2003	12944502
gene therapy for parkinson's disease.	we review recent progress in gene therapy utilizing experimental parkinsonian models including our data. investigation of ex vivo gene therapy for parkinson's disease (pd) is to provide l-dopa by transplantation of genetically modified cells into the striatum. recently, neuronal progenitor cells (npc) are recognized as the most appropriate target population for such genetic and cellular therapy of pd. we have developed modified pseudo-typed retrovirus production system. using this gene transfer ...	2003	12946058
requirements for adeno-associated virus-derived non-viral vectors to achieve stable and site-specific integration of plasmid dna in liver carcinoma cells.	adeno-associated virus (aav) is the only known virus capable of site-specific genomic integration in human cells. thus, aav-based vectors may be an attractive option to achieve prolonged transgene expression in human cells. we therefore studied the minimal elements of gene therapy vectors necessary for stable integration and tested the effectiveness of this approach in hepatoma cells.	2003	12949435
germ-line transmission of a myocardium-specific gfp transgene reveals critical regulatory elements in the cardiac myosin light chain 2 promoter of zebrafish.	in response to the lack of a transgenic line of zebrafish labeled with heart-specific fluorescence in vivo to serve as a research model, we cloned a 1.6-kb polymerase chain reaction (pcr) -product containing the upstream sequence (-870 bp), exon 1 (39 bp), intron 1 (682 bp), and exon 2 (69 bp) of the zebrafish cardiac myosin light chain 2 gene, (cmlc2). a germ-line transmitted zebrafish possessing a green fluorescent heart was generated by injecting this pcr product fused with the green fluoresc ...	2003	12950077
double strand interaction is the predominant pathway for intermolecular recombination of adeno-associated viral genomes.	intermolecular recombination is the foundation for dual vector mediated larger gene transfer by recombinant adeno-associated virus (raav). to identify precursors for intermolecular recombination, we sequentially infected skeletal muscle with aav lacz trans-splicing viruses. at 1 month postinfection, nearly all inputting single-strand (ss) aav genomes were cleared out in muscle. if ss-ss interaction is absolutely required for intermolecular recombination, lacz expression from sequential infection ...	2003	12951015
microdystrophin gene therapy of cardiomyopathy restores dystrophin-glycoprotein complex and improves sarcolemma integrity in the mdx mouse heart.	more than 90% of duchenne muscular dystrophy (dmd) patients develop cardiomyopathy, and many die of cardiac failure. despite tremendous progress in skeletal muscle gene therapy, few attempts have been made to treat cardiomyopathy. microdystrophin genes are shown to correct skeletal muscle pathological lesions in the mdx mouse model for dmd. here, we tested the therapeutic potential of adeno-associated virus (aav)-mediated microdystrophin gene therapy in the mdx mouse heart.	2003	12952841