45 research outputs found

    Using viral vectors as gene transfer tools (Cell Biology and Toxicology Special Issue: ETCS-UK 1 day meeting on genetic manipulation of cells)

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    In recent years, the development of powerful viral gene transfer techniques has greatly facilitated the study of gene function. This review summarises some of the viral delivery systems routinely used to mediate gene transfer into cell lines, primary cell cultures and in whole animal models. The systems described were originally discussed at a 1-day European Tissue Culture Society (ETCS-UK) workshop that was held at University College London on 1st April 2009. Recombinant-deficient viral vectors (viruses that are no longer able to replicate) are used to transduce dividing and post-mitotic cells, and they have been optimised to mediate regulatable, powerful, long-term and cell-specific expression. Hence, viral systems have become very widely used, especially in the field of neurobiology. This review introduces the main categories of viral vectors, focusing on their initial development and highlighting modifications and improvements made since their introduction. In particular, the use of specific promoters to restrict expression, translational enhancers and regulatory elements to boost expression from a single virion and the development of regulatable systems is described

    Molecular studies on chemotactic receptors

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    A block in virus-like particle maturation following assembly of murine leukaemia virus in insect cells

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    Expression of the murine leukaemia virus (MLV) major Gag antigen p65Gag using the baculovirus expression system leads to efficient assembly and release of virus-like particles (VLP) representative of immature MLV. Expression of p180Gag-Pol, facilitated normally in mammalian cells by readthrough of the p65Gag termination codon, also occurs efficiently in insect cells to provide a source of the MLV protease and a pattern of p65Gag processing similar to that observed in mammalian cells. VLP release from p180Gag-Pol-expressing cells however remains essentially immature with disproportionate levels of the uncleaved p65Gag precursor when compared to the intracellular Gag profile. Changing the p65Gag termination codon altered the level of p65Gag and p180Gag-Pol within expressing cells but did not alter the pattern of released VLP, which remained immature. Coexpression of p65Gag with a fixed readthrough p180Gag-Pol also led to only immature VLP release despite high intracellular protease levels. Our data suggest a mechanism that preferentially selects uncleaved p65Gag for the assembly of MLV in this heterologous expression system and implies that, in addition to their relative levels, active sorting of the correct p65Gag and p180Gag-Pol ratios may occur in producer cells

    Lentiviral vector pseudotyped with sendai virus F and HN proteins uses sialylated glycan receptors to efficiently target human airway cells

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    This abstract is for work presented at the 23rd Annual Meeting of the American Society for Gene and Cell Therapy, 12-15 May 2020
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