Viral Vectors in Mammalian Cell Transduction
say.no at spam.now
Sun Jan 18 18:32:02 EST 2004
"LUser <" wrote:
> I am working with a non-continuous line of cultured mammalian cells,
> namely adult bone marrow mesenchymal "stem" cells.
> The big thing for long-term tracking of engrafted cells is to label them
> with reporters like a GFP variant or LacZ.
> Transduction seems to be the best strategy to get the gene in, since we
> want high efficiency of gene delivery in cells that have limited
> population doublings.
> There is no one viral vector to be used. Some use retroviral vectors
> because they want stable transfection (integration)....why? I don't know
> for a cell line that will not be continuous. Others have used adenovirus
> (only transient expression here).
> If I understand correctly, adenovirus seems to make the most sense.
> Infection with replication-defective virions under the right conditions
> is likely to produce higher levels of expression
> (transcription/translation) than use of a retrovirus who integrated
> proviral form may not be a high level expressor. I am led to believe
> that the risk of adenoviral vector labeling is that expression may be
> lost within the time period it is wanted: these grafted stem cells may
> be in the tissues quite a few months before the tissues are examined, and
> the reporter expression must be active to find the cell in the tissue.
> I appreciate any corrections to any misunderstandings I have.
I can't help with your viral transfection questions, but I thought that you
might want to take a look at...
Morris, M.C., Chaloin, L., Mery, J., Heitz, F., and Divita, G. 1999. A novel
potent strategy for gene delivery using a single peptide vector as a
carrier. Nucleic Acids Research, 27(17):3510-3517.
Morris, M.C., Vidal, P., Chaloin, L., Heitz, F., and Divita, G. 1997. A new
peptide vector for efficient delivery of oligonucleotides into mammalian
cells. Nucleic Acids Research, 25(14): 2730-2736.
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