scholarly journals Expression of human cholesterol 7α-hydroxylase in atherosclerosis-susceptible mice via adenovirus infection

1997 ◽  
Vol 324 (3) ◽  
pp. 863-867 ◽  
Author(s):  
Gina L. MOORE ◽  
Christian A. DREVON ◽  
Dietrich MACHLEDER ◽  
John D. TRAWICK ◽  
Alan McCLELLAND ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Transcriptional Repression ◽  
Adenovirus Vector ◽  
Chow Diet ◽  
Adenovirus Infection ◽  
Hepatic Inflammation ◽  
Short Term ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Virus Promoter

Adenovirus is a vector for the delivery of genes mainly to the liver. Short-term (~3 days) studies using adenovirus transfection have provided valuable insights into how genes can complement normal and pathological phenotypes. When atherosclerosis-susceptible C57BL/6 mice were infected with an adenovirus vector containing the human 7α-hydroxylase cDNA (AV17h1) and fed on a chow diet, human 7α-hydroxylase mRNA and enzyme activity doubled compared with that in mice infected with an adenovirus vector (AV1Null) alone. In AV17h1-infected mice fed on a high fat cholic acid (HFCA) diet, mRNA expression and activity of both the endogenous and adenovirus (human) 7α-hydroxylase were repressed. AV17h1-infected mice fed on a HFCA diet and killed at mid-light had increased 7α-hydroxylase activity and mRNA compared with mice killed at mid-dark. Since expression of AV17h1 is driven by a constitutive Rous sarcoma virus promoter, the repression of human 7α-hydroxylase by the HFCA diet was unexpected. In spite of this post-transcriptional repression by the HFCA diet, AV17h1-infected mice expressed the human 7α-hydroxylase mRNA, causing its enzyme activity to be 3-fold greater than in AV1Null-infected mice. In AV17h1-infected mice, the 7α-hydroxylase enzyme activity varied as a linear function of human mRNA abundance. In conclusion, the accumulation of apolipoprotein B-containing lipoproteins in plasma of C57BL/6 mice fed on the HFCA diet was not reduced by longer-term (2 weeks) 7α-hydroxylase expression, probably because of its diminished expression caused by the diet and hepatic inflammation from the adenovirus infection. These results may suggest that adenovirus is effective in promoting longer-term (2 weeks) expression of 7α-hydroxylase.

scholarly journals Generation of an Adenovirus Vector Lacking E1, E2a, E3, and All of E4 except Open Reading Frame 3

1999 ◽  
Vol 73 (7) ◽  
pp. 6048-6055 ◽  
Author(s):  
Mario I. Gorziglia ◽  
Claudia Lapcevich ◽  
Soumitra Roy ◽  
Qiang Kang ◽  
Mike Kadan ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Liver Toxicity ◽  
Adenovirus Vector ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Adenovirus Vectors ◽  
Virus Promoter

ABSTRACT Toxicity and immunity associated with adenovirus backbone gene expression is an important hurdle to overcome for successful gene therapy. Recent efforts to improve adenovirus vectors for in vivo use have focused on the sequential deletion of essential early genes. Adenovirus vectors have been constructed with the E1 gene deleted and with this deletion in combination with an E2a, E2b, or E4 deletion. We report here a novel vector (Av4orf3nBg) lacking E1, E2a, and all of E4 except open reading frame 3 (ORF3) and expressing a β-galactosidase reporter gene. This vector was generated by transfection of a plasmid carrying the full-length vector sequence into A30.S8 cells that express E1 and E2a but not E4. Production was subsequently performed in an E1-, E2a-, and E4-complementing cell line. We demonstrated with C57BL/6 mice that the Av4orf3nBg vector effected gene transfer with an efficiency comparable to that of the Av3nBg (wild-type E4) vector but that the former exhibited a higher level of β-galactosidase expression. This observation suggests that E4 ORF3 alone is able to enhance RNA levels from the β-galactosidase gene when the Rous sarcoma virus promoter is used to drive transgene expression in the mouse liver. In addition, we observed less liver toxicity in mice injected with the Av4orf3nBg vector than those injected with the Av3nBg vector at a comparable DNA copy number per cell. This study suggests that the additional deletion of E4 in an E1 and E2a deletion background may be beneficial in decreasing immunogenicity and improving safety and toxicity profiles, as well as increasing transgene capacity and expression for liver-directed gene therapy.

scholarly journals Comparison of the relative importance of tyrosine-specific vinculin phosphorylation and the loss of surface-associated fibronectin in the morphology of cells transformed by Rous sarcoma virus

1986 ◽  
Vol 82 (1) ◽  
pp. 129-142
Author(s):  
S. Kellie ◽  
B. Patel ◽  
A. Mitchell ◽  
D.R. Critchley ◽  
N.M. Wigglesworth ◽  
...  
Keyword(s):  
Rous Sarcoma Virus ◽  
Relative Importance ◽  
Plasma Fibronectin ◽  
Short Term ◽  
Normal Morphology ◽  
Rous Sarcoma ◽  
Microfilament Bundles ◽  
Sarcoma Virus ◽  
Adhesion Plaques

We have investigated the relative importance of tyrosine-specific phosphorylation of vinculin and the loss of surface-associated fibronectin in the maintenance of the rounded morphology characteristic of chick embryo fibroblasts (CEF) transformed by Rous sarcoma virus (RSV). To address this question we have examined the interaction of CEF and RSV-CEF in vitro with exogenously added fibronectin in both 3-day culture experiments and short-term, 3-h spreading experiments. We report that the addition of human plasma fibronectin to cultures of RSV-CEF results in the restoration of a near-normal morphology, as has been described previously, with the added fibronectin incorporated into an extracellular matrix. However, the phosphotyrosine content of vinculin in these cells was unchanged from that of control, untreated RSV-CEF despite the change in morphology. In short-term spreading experiments RSV-CEF were unable to adopt a fully spread morphology on fibronectin substrates, with defects in the formation of adhesion plaques and microfilament bundles compared with untransformed CEF. pp60v-src was present in the newly formed adhesion plaques of RSV-CEF spreading on fibronectin substrates. The relevance of these results to the maintenance of the transformed phenotype is discussed.

Vincristine-resistant human laryngeal carcinoma cells demonstrate increased Rous sarcoma virus promoter activity

Life Sciences ◽  
2010 ◽  
Vol 87 (15-16) ◽  
pp. 468-474 ◽  
Author(s):  
Dragomira Majhen ◽  
Anamaria Brozovic ◽  
Tvrtko Buger ◽  
Jelka Gabrilovac ◽  
Maja Osmak ◽  
...  
Keyword(s):  
Laryngeal Carcinoma ◽  
Rous Sarcoma Virus ◽  
Promoter Activity ◽  
Carcinoma Cells ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Human Laryngeal Carcinoma ◽  
Virus Promoter

Reactivation of Transgene Expression by Alleviating CpG Methylation of the Rous sarcoma virus Promoter in Transgenic Quail Cells

2011 ◽  
Vol 49 (3) ◽  
pp. 222-228 ◽  
Author(s):  
Hyun-Jun Jang ◽  
Jin Won Choi ◽  
Young Min Kim ◽  
Sang Su Shin ◽  
Kichoon Lee ◽  
...  
Keyword(s):  
Rous Sarcoma Virus ◽  
Transgene Expression ◽  
Cpg Methylation ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Virus Promoter

scholarly journals Sense and antisense modification of glial alpha B-crystallin production results in alterations of stress fiber formation and thermoresistance.

1994 ◽  
Vol 125 (6) ◽  
pp. 1385-1393 ◽  
Author(s):  
T Iwaki ◽  
A Iwaki ◽  
J Tateishi ◽  
J E Goldman
Keyword(s):  
Glioma Cells ◽  
Fiber Formation ◽  
C6 Glioma Cells ◽  
Cytoskeletal Organization ◽  
Stable Transfectants ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
The Stability ◽  
Virus Promoter ◽  
Rat C6 Glioma

The phenotypic effects of selectively altering the levels of alpha B-crystallin in cultured glial cells were analyzed using sense and antisense approaches. Rat C6 glioma cells and human U-373MG glioma cells were transfected with a rat alpha B-crystallin sense cDNA or an antisense cDNA regulated by a Rous sarcoma virus promoter to alter cellular levels of alpha B-crystallin. The antisense strategy resulted in decreased alpha B-crystallin levels, as revealed by Western blot and immunocytochemical analyses. The reduced alpha B-crystallin expression was accompanied by alterations in cellular phenotype: (a) a reduction of cell size and/or a slender cell morphology; (b) a disorganized microfilament network; and (c) a reduction of cell adhesiveness. Like HSP27, the presence of additional alpha B-crystallin protein confers a thermoresistant phenotype to stable transfectants. Thus, alpha B-crystallin in glioma cells plays a role in their thermal resistance and may contribute to the stability of cytoskeletal organization.

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