plant viruses as gene transfer vectors

... of plant genes was established as AACAATGG (8) A recent comparison of the effect of these two consensus sequences placed upstream of the P-glucuronidase gene (gus) in plant protoplasts has Genes ... Chloramphenicol Acetyltransferase Gene The most commonly used chloramphenicol acetyltransferase gene (cat) originates from transposon Tn9 (20) It has been widely used as a reporter gene in mammalian ... phosphinothricin acetyltransferase (bar) genes can also be quantified using radiochemtcal assays (22,23) Selectable Marker Genes A selectable marker gene is used to recover transformants after a gene transfer...

Ngày tải lên: 11/04/2014, 10:10

... of transgene expression for at least months [28], providing increased optimism for the use of gene transfer in the treatment of chronic articular disease Indeed, IL-1Ra gene therapy has demonstrated ... of gene transfer as a method of drug delivery was perhaps most effectively illustrated The maintenance of and gradual increase in IL-1Ra concentration provided by ongoing synthesis by the genetically ... was shifted slightly to the left, indicating a slight increase in the effectiveness of IL-1Ra when provided as a transgene product This may arise from increased concentrations of the IL-1Ra gene...

Ngày tải lên: 09/08/2014, 01:23

... codon-optimized lacZ gene sequence encoding β-galactosidase (βGal) [29] was used to replace the EGFP transgene sequence The VSV-G-encoding pLTR-G plasmid was described before [30] Plasmids encoding ... pseudotyped HIV1-based gene transfer vectors Gene Therapy 2000, 7:910-913 Marino MP, Luce MJ, Reiser J: Small- to large-scale production of lentivirus vectors In Lentivirus Gene Engineering Protocols ... nervous system diseases Curr Gene Ther 2007, 7:239-248 Kato S, Inoue K, Kobayashi K, Yasoshima Y, Miyachi S, Inoue S, Hanawa H, Shimada T, Takada M, Kobayashi K: Efficient gene transfer via retrograde...

Ngày tải lên: 14/08/2014, 19:22

... mosaicism associated with these vectors However, the new generation of lentiviral vectors would avoid such problems [21, 22, 23] These vectors have the additional advantage of high gene transfer ... with the embryo-based gene transfer methods considered above However, there are at least some glimmers of hope for future gene targeting possibilities in the case of embryo-based approaches: ... episomal plasmid vectors tend to behave in an unstable fashion [76, 77] During development, plasmid copy numbers fluctuate and plasmids are lost from some cells Plasmid inheritance to subsequent generations...

Ngày tải lên: 03/11/2012, 10:01

... that was absent in cytoplasmic fractions (Fig 3Ai) Further, most of the cytochrome C was confined to mitochondrial rich fractions and very little of cytochrome C was released into the cytoplasmic ... majority of the overexpressed MCD was the short form and was localized to mitochondria, the long form was expressed in Ad.MCD cells and was observed primarily in cytoplasm As mitochondria are a rich ... MCD was present in both cytoplasmic and mitochondrial rich fractions While the majority of over expressed MCD activity was present in mitochondria, about 30–40% of total MCD activity was measured...

Ngày tải lên: 16/03/2014, 16:20

... been tested in phase I clinical trials with impressive safety data Moreover, successful gene transfer has been demonstrated in most cases in which it has been analyzed In contrast, only rare examples ... p53 gene transfer to ovarian cancer cells using cationic nonviral vector has also been reported [21] Adp53 was evaluated in a phase I/II trial and the treatment was well tolerated [9,22,23] Gene ... Beta-galactosidase gene transfer to human malignant glioma in vivo using replication-deficient retroviruses and adenoviruses Hum Gene Ther 9: 1769 – 1774 86 Kanerva, A., et al (2005) Noninvasive dual...

Ngày tải lên: 22/03/2014, 11:20

... Additionally, in sulfatase-deficient LSDs, the coexpression of a sulfatase gene together with the sulfatase-modifying factor Promoter and SUMF1 effect on Morquio gene transfer (SUMF1) gene has permitted ... transfer was performed as described previously [85] One unit was defined as the catalysis of nmol of substrate h)1 GALNS activity was expressed as UÆmL)1 (medium) or UÆmg)1 protein (cell lysate), as ... effect on Morquio gene transfer 48 Vargas J Jr, Gusella GL, Najfeld V, Klotman ME & Cara A (2004) Novel integrase-defective lentiviral episomal vectors for gene transfer Hum Gene Ther 15, 361–372...

Ngày tải lên: 29/03/2014, 21:20

... Engelhardt Introduction Adeno-associated virus (AAV) as a vector for gene therapy of cardiovascular diseases has received much recent attention (1–6) This enthusiasm is based on the success of numerous ... followed by an assay for the reporter gene product We typically include a β-galactosidase gene in our gutted viruses and a human alkaline phosphatase gene in the helper viruses, both under the control ... Adenoviral Vectors 19 Gutted Adenoviral Vectors for Gene Transfer to Muscle Jeannine M Scott and Jeffrey S Chamberlain Introduction Adenoviral vectors are a popular choice for gene transfer protocols...

Ngày tải lên: 10/04/2014, 11:11

... (DEAE-dextran), with a plasmid vector that has regulatory sequences adjacent to the chloramphenicol acetyl transferase (CAT) gene This type of gene transfer is often referred to as a transient expression ... polybrene/DMSO-assisted gene transfer as optimized for NIH 3T3 fibroblasts polybrene/DMSO-assisted gene transfer can be performed by simply incorporating the appropriate set of conditions into the basic transfection ... pSV2neo plasmid DNA (seeNote 5) A schematic representation of the basic polybrene/DMSO-assisted transfer protocol is provided in Fig gene- 3.2 Optimized Parameters for PolybrenelDMSO-Assisted Gene...

Ngày tải lên: 11/04/2014, 09:42

... there has been remarkably rapid progress in defining the molecular basis of inherited disorders Many disease genes (the majority of which are genes responsible for monogenic Mendelian diseases) ... complex diseases, such as hypertension, diabetes mellitus, and coronary artery diseases, each of which involves many susceptibility genes Disease-causing mutations can be broadly classified into ... Analysis of Genetic Variants in Cardiovascular Risk Genes by Heteroduplex Analysis Ana Cenarro, Fernando Civeira, and Miguel Pocovi Introduction As an increasing number of human diseases are linked...

Ngày tải lên: 11/04/2014, 10:39

... normalized against actin DNA in each sample Gene transfer assay Gene transfer efficacy of the viral vectors was assessed with the use of GFP reporter In the assay, cells were plated in 24-well plates ... Competitive inhibition assay showing the enhanced gene transfer efficacy of Ad5 Figure Competitive inhibition assay showing the enhanced gene transfer efficacy of Ad5.PTDtat was mediated by the PTDtat ... exhibited enhanced gene transfer efficacy in a variety of tumor cells Ad5.PTD Ad5.PTDtat exhibited enhanced gene transfer efficacy in a variety of tumor cells Gene transfer efficacy was evaluated by...

Ngày tải lên: 18/06/2014, 18:20

... +-ATPase modulates cardiac contraction and relaxation Cardiovasc Res 2003, 57:20-27 Kawase Y, Hajjar RJ: The cardiac sarcoplasmic/endoplasmic reticulum calcium ATPase: a potent target for cardiovascular ... target for cardiovascular diseases Nat Clin Pract Cardiovasc Med 2008, 5:554-565 Lipskaia L, Chemaly ER, Hadri L, Lompre AM, Hajjar RJ: Sarcoplasmic reticulum Ca(2+) ATPase as a therapeutic target ... 17:253-261 26 Hadri L, Bobe R, Kawase Y, Ladage D, Ishikawa K, Atassi F, Lebeche D, Kranias EG, Leopold JA, Lompre AM, Lompré AM, Lipskaia L, Hajjar RJ: SERCA2a gene transfer enhances eNOS expression...

Ngày tải lên: 18/06/2014, 22:20

... normalized against actin DNA in each sample Gene transfer assay Gene transfer efficacy of the viral vectors was assessed with the use of GFP reporter In the assay, cells were plated in 24-well plates ... Competitive inhibition assay showing the enhanced gene transfer efficacy of Ad5 Figure Competitive inhibition assay showing the enhanced gene transfer efficacy of Ad5.PTDtat was mediated by the PTDtat ... exhibited enhanced gene transfer efficacy in a variety of tumor cells Ad5.PTD Ad5.PTDtat exhibited enhanced gene transfer efficacy in a variety of tumor cells Gene transfer efficacy was evaluated by...

Ngày tải lên: 20/06/2014, 01:20

... +-ATPase modulates cardiac contraction and relaxation Cardiovasc Res 2003, 57:20-27 Kawase Y, Hajjar RJ: The cardiac sarcoplasmic/endoplasmic reticulum calcium ATPase: a potent target for cardiovascular ... target for cardiovascular diseases Nat Clin Pract Cardiovasc Med 2008, 5:554-565 Lipskaia L, Chemaly ER, Hadri L, Lompre AM, Hajjar RJ: Sarcoplasmic reticulum Ca(2+) ATPase as a therapeutic target ... 17:253-261 26 Hadri L, Bobe R, Kawase Y, Ladage D, Ishikawa K, Atassi F, Lebeche D, Kranias EG, Leopold JA, Lompre AM, Lompré AM, Lipskaia L, Hajjar RJ: SERCA2a gene transfer enhances eNOS expression...

Ngày tải lên: 20/06/2014, 04:20

... as well as able to block inflammation in vivo, it has been proposed that local intra-articular gene transfer of TGF-β1 could be therapeutic for the treatment of rheumatoid arthritis as well as ... there was evi- dence of both fibroblast and myofibroblast proliferation between myofibers with intracellular edema, but there was no evidence of inflammation or myonecrosis (Fig 5M,N) In contrast, ... Ad.TGF-β1 in the knee joints of naïve C57/Bl/6 mice where gene transfer of TGF-β1 to the mouse knee resulted in hyperplasia of the synovium as well as in chondro-osteophyte formation at the chondrosynovial...

Ngày tải lên: 09/08/2014, 01:21

... Local gene transfer of IL-13 The recombinant adenovirus encoding human IL-13 (AxCAhIL-13) was generated as described before [17-19] and an empty adenoviral construct (AxCANI) was used as control ... dehydrogenase; MMP, matrix metalloproteinase Three sections of each knee joint were measured and the mean was calculated We report the amount of positive features per 100,000 µm2 inflammatory cell mass ... expression by 43%, as only 20% VDIPEN expression was found in the IL-13 group The inhibitory effect of IL-13 was still present at day after arthritis induction, as only 10% VDIPEN expression was found...

Ngày tải lên: 09/08/2014, 06:22

... C, Boissier MC: Syngeneic fibroblasts transfected with a plasmid encoding interleukin-4 as non-viral vectors for antiinflammatory gene therapy in collagen-induced arthritis J Gene Med 2002, 4:300-307 ... direct viral gene transfer of therapeutic agents provide proof supporting the use of gene therapy in arthritis [9-13,32,33], and a recent phase I clinical trial using IL-1Ra by ex vivo gene transfer ... arthritic joints was also successfully completed [40,41] Systemic non-viral delivery methods, such as administration of engineered syngeneic fibroblasts [42], and intranasal delivery of plasmid DNA [43]...

Ngày tải lên: 09/08/2014, 08:22

... Conclusions Adenoviral BMP-4 gene transfer efficiently induces the chondrogenic differentiation of human primary MSCs as effectively as BMP-2 gene transfer However, both transgenes induced high levels ... recombinant adenoviral vectors The complete coding sequence of the human BMP-4 gene [GenBank:M22490] cloned into λ gt10 bacteriophage vectors (ATCC No 40342; Manassas, VA, USA) was isolated and purified ... serotype adenoviral vectors carrying the cDNAs for human BMP-4 were constructed by cre-lox recombination as previously described [22] The vectors encoding BMP-2, firefly luciferase (Luc) or green...

Ngày tải lên: 09/08/2014, 14:22

... for HGT The over-hasty assignment of recent bacterial-to-vertebrate gene transfers, solely on the basis of BLAST E-values [31], has been firmly refuted [32,33] Such premature HGT assignments have ... luminescens Pseudomonas syringae Pseudomonas putida Pseudomonas putida Bordetella pertussis Bordetella pertussis Xylella fastidiosa 0.95 Xanthomonas axonopodis Xylella fastidiosa Xanthomonas axonopodis ... The TEP genes have been amplified by a process of tandem duplication into linked multigene families Drosophila melanogaster has six TEP genes, whereas the mosquito Anopheles gambiae has 15 [13]...

Ngày tải lên: 09/08/2014, 20:20

... resistance gene and a SV40 polyadenylation signal containing cassette was cloned at the EcoRV site to derive the control RFP SB plasmid To generate this pIRESneoRFP cassette, RFP gene was cloned as a ... shRNA DNA cassette was cloned in parallel as a BglII-BglII fragment in the pT/BH plasmid at BamHI site to get pT/BH-U6CXCR4 or pT/BH-H1CCR5 The CMV-RFP-IRES-neo-SV40pA cassette was released as NruI-BstZ17I ... either conventional retroviral vectors or lentiviral vectors were used for gene transfer Although highly efficient for stable gene transfer, use of retroviral derived vectors poses a degree of risk...

Ngày tải lên: 10/08/2014, 05:21