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.
> 

Kevin Merrell




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