in vitro Knock out

Robert M. Mihalek mihalek at FORMULA1.smtp.anes.upmc.edu
Mon Aug 23 09:22:09 EST 1999


In article <37C01FF5.5097B353 at pasteur.fr>
Pierre Rodrigues <pirod at pasteur.fr> writes:

> Hello,
> 
> I was wondering if there is a way to perform a gene knock out in a cell
> culture without going on the mouse. Just to make an homologous
> recombination and select resistant clones which would be knocked out in
> a gene...
> 
> Is there anybody out there which has a clue?
> 
> thanks
> 
> Pierre Rodrigues
> Institut Pasteur
> France

It may be possible to knock-out both copies of the gene-of-interest by
doing successive electroporations, perhaps using different targeting
vectors for each so that you can select for a different antibiotic
resistance.

I have heard that the amount of manipulations required for multiple
targeting events can leave the ES cells very damaged. 


Bob Mihalek=================================================
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