gene targeting in cell type other than ES
Kevin T. Merrell
ktm3 at columbia.edu
Sun Jan 8 13:16:49 EST 1995
On Thu, 5 Jan 1995, Riccardo Fodde wrote:
> Hi everybody,
> I would like to ask for information on gene targeting in EBV-transformed
> celllines. Has anybody tried this and how to get single colonies?
We do this with v-abl transformed mouse pre-B cells. We obtain clones by
plating the electroporated cells in 96 well plates at a low enought
dilution that only a portion of the wells end up with resistant cells and
are thus likely to represent a clonal population. Don't forget, for any
autosomal gene you may need to knock-out both copies.
> Also I would like to know if it is possible to target primary fibroblasts
> without having to immortalize them first and still have enough passages left
> to do something with correctly targeted cells.
I doubt it.
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