I want to produce stable cell lines of Huh7 and Huh7D which express functional TLR3.
I would like to label this TLR3 with BFP in order to save myself some time when It comes time to select colonies.
I have made stable huh7.5 cell lines before, which express fluorescently labeled IPS1, so I am familiar with the process.
However, I do not want to affect TLR3 function and I don't want BFP to be cleaved (obvious concerns)
So, my question is: If possible, what terminus of TLR3 should I add BFP to?
Cindy Körner
Why don't you just use a construct where the fluorescent marker is not fused to your TLR but is expressed with an IRES. You still have your positive clones and don't have to be afraid that it might interfere with TLR function.
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