Restoring the FLP site in HEK293T Flp-In cells

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Restoring the FLP site in HEK293T Flp-In cells

Postby HEK293T » Mar 21 2017 1:53 pm

Hi all,

Just wanted to quickly clarify something: If I have a Flp-In cell line that's already expressing my inserted gene in the Flp site and I transfect with pOG44 alone, will my gene be 'flipped' back out restoring the Flp site? Can I then select again with Zeocin? And then retransfect the surviving cells with another pcDNA5 vector to generate a cell line expressing a different gene?

For context, my plan is to do this with a cell line that currently has Cas9 expressed from the FLP site. After generating a knockout, I'd then like to remove Cas9 and have the FLP site available for complementation.

Thanks in advance for your help!
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