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adenoviral cre or lentiviral cre for mouse primary cells?


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#1 tonli

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Posted 07 April 2012 - 08:45 PM

Which one has less cytotoxicity for mouse primary cells, like MEF? Thanks a lot!

#2 doxorubicin

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Posted 09 April 2012 - 11:35 AM

In my experience, you can find a concentration (MOI) of adenovirus expressing cre that gives you almost 100% recombination with minimal cell death. Too much virus will give you more death, and less virus will live you less recombination...you just need to find the right "dose". Any time I used lentivirus, I infected a very small percentage of cells and then had to select infected cells by drug resistance or GFP expression. This is difficult to do in primary MEFs, since the cells will encounter replicative senescence and die before you have enough cells to work with.

#3 tonli

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Posted 09 April 2012 - 07:55 PM

I see, thanks a lot for sharing!

In my experience, you can find a concentration (MOI) of adenovirus expressing cre that gives you almost 100% recombination with minimal cell death. Too much virus will give you more death, and less virus will live you less recombination...you just need to find the right "dose". Any time I used lentivirus, I infected a very small percentage of cells and then had to select infected cells by drug resistance or GFP expression. This is difficult to do in primary MEFs, since the cells will encounter replicative senescence and die before you have enough cells to work with.






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