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Production of high titer helper-free retroviruses by transient transfection

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TLDR
In this article, a method for producing high-titer, helper-free infectious retroviruses is disclosed which employs a novel strategy that uses transient transfection of new retroviral producer cell lines, ecotropic line BOSC 23 and amphotropic line CAK 8.
Abstract
A method for producing high-titer, helper-free infectious retroviruses is disclosed which employs a novel strategy that uses transient transfection of new retroviral producer cell lines, ecotropic line BOSC 23 and amphotropic line CAK 8, both of which cell lines and their precursor cell lines are disclosed. Because of the advantages over stable packaging cell lines, the BOSC 23 and CAK 8 transient transfection systems greatly facilitate and extend the use of helper-free retroviral vectors. The cell lines and corresponding methods possess wide application in both the medical and biotechnical fields, including gene therapy. These potential applications are disclosed and illustrated.

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TCR and Lat are expressed on separate protein islands on T cell membranes and concatenate during activation

TL;DR: The spatio-temporal dynamics of T cell antigen receptor (TCR) complexes and linker for activation of T cells (Lat), a key adaptor molecule in the TCR signaling pathway, are analyzed in T cell membranes using high-speed photoactivated localization microscopy, dual-color fluorescence cross-correlation spectroscopy and transmission electron microscopy.
Journal ArticleDOI

Biochemical Mechanisms of IL-2–Regulated Fas-Mediated T Cell Apoptosis

TL;DR: The ability of IL-2 to enhance expression of a pro-apoptotic molecule, FasL, and to suppress an inhibitor of Fas signaling, FLIP, likely accounts for the role of this cytokine in potentiating T cell apoptosis.
Journal ArticleDOI

Endothelial adherens junctions control tight junctions by VE-cadherin-mediated upregulation of claudin-5.

TL;DR: It is shown that endothelial VE-cadherin at AJs upregulates the gene encoding the TJ adhesive protein claudin-5, and this effect requires the release of the inhibitory activity of forkhead box factor FoxO1 and the Tcf-4–β-catenin transcriptional repressor complex.
Journal ArticleDOI

A role for Rac in Tiam1-induced membrane ruffling and invasion.

TL;DR: In fibroblasts, Tiaml induces a similar phenotype as constitutively activated (Vll)Racl, including membrane ruffling, and this is inhibited by dominant negative (N17)Racan, indicating that the TiamL-Rae signalling pathway could be operating in the invasion and metastasis of tumour cells.
Journal ArticleDOI

SIRT2 deacetylates FOXO3a in response to oxidative stress and caloric restriction

TL;DR: It is reported here that the expression of SIRT2 is elevated in the white adipose tissue and kidney of caloric‐restricted mice and it is demonstrated that SIRT3a binds to FOXO3a and reduces its acetylation level, which increases FOXO DNA binding and elevates theexpression of FOXO target genes.
References
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Journal ArticleDOI

Characteristics of a Human Cell Line Transformed by DNA from Human Adenovirus Type 5

TL;DR: Human embryonic kidney cells have been transformed by exposing cells to sheared fragments of adenovirus type 5 DNA, and the transformed cells exhibited many of the characteristics of transformation including the elaboration of a virus-specific tumour antigen.
Journal ArticleDOI

Vaccination with Irradiated Tumor Cells Engineered to Secrete Murine Granulocyte-Macrophage Colony-Stimulating Factor Stimulates Potent, Specific, and Long-Lasting Anti-Tumor Immunity

TL;DR: The results have important implications for the clinical use of genetically modified tumor cells as therapeutic cancer vaccines and the levels of anti-tumor immunity reported previously in cytokine gene transfer studies involving live, transduced cells could be achieved through the use of irradiated cells alone.
Journal ArticleDOI

Induction of chronic myelogenous leukemia in mice by the P210bcr/abl gene of the Philadelphia chromosome

TL;DR: It is demonstrated that P210bcr/abl expression can induce chronic myelogenous leukemia and retrovirus-mediated expression of the protein provides a murine model system for further analysis of the disease.
Journal ArticleDOI

The Basic Science of Gene Therapy

TL;DR: A large number of key technical issues need to be resolved before gene therapy can be safely and effectively applied in the clinic, and future technological developments will be critical for the successful practice of gene therapy.
Journal Article

Improved Retroviral Vectors for Gene Transfer and Expression

TL;DR: A set of murine retrovirus-based vectors that include unique cloning sites for insertion of cDNAs such that the cDNA can be driven by either the retroviral long terminal repeat, the immediate early promoter of human cytomegalovirus, or the simian virus 40 early promoter are described.
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