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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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Development of AAVLP(HPV16/31L2) Particles as Broadly Protective HPV Vaccine Candidate

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Calcium-phosphate mediated DNA transfer into HEK-293 cells in suspension: control of physicochemical parameters allows transfection in stirred media. Transfection and protein expression in mammalian cells

TL;DR: This is the first report of transient transfection of suspended cells with purified plasmid DNA in bioreactors or spinner flasks of the immortalized human embryo kidney cell line 293, and the protocol for the generation of precipitate complexes was modified based on earlier work.
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Foxp3 Protein Stability Is Regulated by Cyclin-dependent Kinase 2

TL;DR: The results indicate that CDK2 negatively regulates the stability and activity of Foxp3 and implicate CDK-coupled receptor signal transduction in the control of regulatory T cell function and stability.
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Regulation of the Oncogenic Activity of BCR-ABL by a Tightly Bound Substrate Protein RIN1

TL;DR: RIN1 rescues BCR-ABL mutants for transformation in a manner distinguishable from the cell cycle regulators c-Myc and cyclin D1 and the Ras connector Shc and exemplifies a new class of effector molecules dependent on the concerted action of the SH3, SH2, and catalytic domains of a cytoplasmic tyrosine kinase.
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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