A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5

McGrory, W.J.; Bautista, Diosdado S.; Graham, Frank L.

doi:10.1016/0042-6822(88)90302-9

Cited by 575 publications

(311 citation statements)

References 19 publications

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“…8 The resulting plasmid (pAV-rtTA) was cotransfected with pJM17 into 293 cells via calcium phosphate precipitation and the adenovirus plaque purified, amplified and titrated by standard techniques. 17,18 Rat 9L gliosarcoma cells (a gift of E Oldfield, NINDS, NIH) were plated at a density of 1 × 10 6 in T75 flasks in Dulbecco's modified Eagle's medium (DMEM; Life Technologies, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS) and 50.0 g/ml gentamicin sulfate and grown overnight. Adenoviruses were added to target cells in a minimal volume of medium at the MOIs shown in Table 1.…”

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confidence: 99%

Adeno-retroviral chimeric viruses as in vivo transducing agents

et al. 1999

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Several hybrid viral gene transfer systems have been all four transcomplementing adeno-retroviruses. Molecular described that exploit the favorable features of the two paranalysis of DNA extracted from 9L G418 R populations ent viral species. We have developed a hybrid adeno-retroderived both in vitro and in vivo showed appropriate inteviral vector system to generate a retroviral vector in situ.gration of the LXSN proviral sequence. Tumor cells were The system consists of adenoviruses encoding MoMLV harvested 1, 3 and 4 weeks after adeno-retrovirus adminisgag.pol (Axtet.gag.pol), the VSV-G viral envelope tration to the human A375 xenografts. The percentage of (Axtet.VSV-G), the retroviral vector LXSN expressing the G418 R colonies recovered from tumors transduced with all neomycin phosphotransferase gene (AV-LXSN) and a of the transcomplementing adeno-retroviruses increased transcriptional regulator to control expression of gag.pol with time, whereas no increase was observed in tumors and envelope (AV-rtTA). In vitro, biologically active retrovitransduced with AV-LXSN alone. DNA extracted from ral vector preparations were generated following adeno-G418 R A375 cell populations showed the presence of interetroviral transduction of 9L rat glioma cells. In vivo the grated proviral sequences only in animals that received the transcomplementing adeno-retroviruses were co-adminisfull complement of adeno-retroviruses. These results demtered intratumorally into subcutaneous 9L glioma tumors in onstrate that adenoviral vectors expressing transcomprats and human A375 melanoma xenografts in nude mice.lementing genes for retroviral proteins and retroviral vector In the 9L rat model, G418 R cell cultures were only obtained RNAs can be used for in situ transduction of target cells. when 9L cells were harvested from tumors injected with Keywords: adenovirus; retrovirus; gene therapy There have been several recent reports describing the use of recombinant viruses of one species to provide transcomplementing functions which support the amplification of replication-deficient variants of a second viral species. 1-8 These reports have primarily been in vitro studies which demonstrate the principle that virusmediated provision of these transcomplementing functions is adequate to support the generation of a second vector. Feng and co-workers 7 although have reported evidence that this can occur in vivo using an adeno-retroviral hybrid system. 7 The system, consisting of two adenoviruses, one carrying a retroviral transcription unit encoding green fluorescent protein (GFP) and a second expressing retroviral gag.pol and env functions, was used to transduce human ovarian tumor cells (SKOV3. p1 ) in vivo. Co-injection of both adenoviruses resulted in more efficient transduction of the tumor compared with administration of the adenovirus carrying the retroviral GFP vector alone. To examine further the potential appli-

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confidence: 99%

Adeno-retroviral chimeric viruses as in vivo transducing agents

et al. 1999

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“…After cotransfection of the transfer vector 18 conTherefore, a careful optimization of the conditions used is of great importance to achieve optimal transduction taining C3 with the plasmid pJM17 19 into 293 cells, fluorescence originating from the expression of the nonrecomefficiencies. For those purposes, several adenoviral vectors have been constructed that express marker genes bined plasmid could already be detected in the transfected cells after 16 h, allowing direct determination such as ␤-galactosidase and luciferase that are detected by relatively simple enzymatic assays.…”

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Adenovirus-mediated expression of green fluorescent protein

et al. 1997

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A recombinant replication-deficient adenovirus has been blotting. As demonstrated by FACS analysis, up to 98% of generated that expresses a mutant of the Aquorea victoria HUVEC and approximately 70% of human smooth muscle green fluorescent protein (GFP) under the control of the cells could be transduced to express GFP. Since GFP can strong CMV promoter by insertion into the E1 region (AdVbe detected in cells without the need for prior fixing and GFP). High expression of GFP was found in different cell staining, the virus should be useful for optimizing in living types after infection with the recombinant virus that could cells the transduction efficiency of different cell types, of be easily detected by fluorescence microscopy. In human cells from different experimental animals, as well as studyumbilical vein endothelial cells (HUVEC), expression levels ing the kinetics and persistence of adenovirus-mediated had already reached a maximum after 2 days and were gene transfer in diverse experimental settings. stable for at least 7 days, as determined by Western Keywords: green fluorescent protein; recombinant adenovirus; marker gene; endothelium Gene transfer techniques using replication-deficient of them can be directly visualized in living cells without the need for prior incubation with substrates or staining recombinant adenoviral vectors (AdV) have considerable potential both at experimental and therapeutic levels. [1][2][3] procedures, a feature fulfilled by green fluorescent protein. Advantages include the ability to produce high-titer stocks, the high efficiency of gene transfer into a variety Originally isolated from the jellyfish Aquorea victoria as a 238 amino acid protein, 15 wild-type GFP emits a bright of cell types, and the ability to transduce cells that have a low mitotic index, eg the endothelium, kidney and cells green fluorescence upon excitation at both 395 and 475 nm. Crystallization has revealed a cylinder-like structure of the central nervous system. 4,5 Limitations of AdV include the relatively short time of expression of the of ␤-sheets wrapped around a single central helix. 16 The fluorophore results from the autocatalytic cyclization (therapeutic) gene in vivo (a few weeks to several months, depending on the organ) due to the episomal maintebetween amino acids of the polypeptide backbone and oxidation of the ␣-␤ bond of the central tyrosine. Due to nance of the adenoviral DNA in the cell, and the elicitation of an immune response by the host organism. [6][7][8] this rigid encapsulation, the fluorophore is remarkably insensitive to changes in environmental conditions, eg These features favor, in the first instance, applications where only a transient expression of the ectopic gene is pH, O 2 quenching of the excited state, and to proteolysis. The generation of mutants that show improved levels of desirable, eg inflammatory reactions.Despite its broad cell-type specificity, considerable diffluorescence as well as different excitation and emission maxima has further triggered the widesprea...

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“…We also constructed a type 5 adenoviral vector harboring the bgalactosidase gene (Ad:LacZ) to examine transduction efficiency and as a control. For homologous recombination, a shuttle vector, pAd-YC2, and a rescue vector, pJM17, 46,47 were co-transfected into 293 cells with Tfx20t (Promega, Madison, WI, USA) according to the manufacturer's protocol. In all, 293 cells were plated 24 h prior to transfection at 70% confluency in a 24-well plate.…”

Section: Recombinant Ad Vectorsmentioning

confidence: 99%

Suppression of glomerulosclerosis by adenovirus-mediated IL-10 expression in the kidney

Kim

et al. 2003

Gene Ther

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A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5

Cited by 575 publications

References 19 publications

Adeno-retroviral chimeric viruses as in vivo transducing agents

Adeno-retroviral chimeric viruses as in vivo transducing agents

Adenovirus-mediated expression of green fluorescent protein

Suppression of glomerulosclerosis by adenovirus-mediated IL-10 expression in the kidney

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