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    High Frequency Cell Surface Expression Of A Foreign Gene Product After Murine Hematopoietic Stem Cell Transduction With A Retroviral Vector

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    1996_WilliamsC_THS_000332.pdf (3.995Mb)
    Author
    Williams, Christopher
    Advisor
    Marilyn Schendel; John Addis; Daniel Tumas
    Date of Issue
    1996-04-01
    Metadata
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    URI
    https://scholars.carroll.edu/handle/20.500.12647/2893
    Title
    High Frequency Cell Surface Expression Of A Foreign Gene Product After Murine Hematopoietic Stem Cell Transduction With A Retroviral Vector
    Type
    thesis
    Abstract
    A retroviral vector (pSFF) derived from murine Friend spleen focus forming virus was used to transduce murine hematopoietic stem cells and express a cell surface marker protein, mutated murine prion protein, in vivo after transplantation. To enhance retroviral vector integration in bone marrow cells, mice were treated with 5 - fluorouracil (5- FU) to increase stem cell mitotic activity. The infectivity titer of the vector, pSFF-mPrP-3F4, was determined by a novel assay in which antigen-positive foci of infected cells were detected after replication and spread of the vector in cultures of mixed packaging cell lines. Infection of Sca- 1 + /Lineagene9''low cells with pSFF-mPrP-3F4 resulted in marker protein expression in 40% of the progeny cells after 7 days of culture. Transplantation of marrow cells or sorted Sca- 1 + /Lineagene9'low cells transduced with vector resulted in 3F4- positive mPrP expression in 11-37% of donor-derived peripheral blood leukocytes at two weeks. Although the percentage of 3F4-positive blood cells gradually declined, at 28 weeks 23% of recipient mice still maintained expression of the marker gene. Expression was observed in lymphoid, myeloid and erythroid lineages and was detected in Scal + /Lineagene9_low marrow cells. The multi-lineage, high frequency expression observed suggests that pSFF may have utility in gene therapy directed to hematopoietic stem cells and their differentiated progeny.
    Degree Awarded
    Bachelor's
    Semester
    Spring
    Department
    Life & Environmental Sciences
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    • Life and Environmental Sciences Undergraduate Theses

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