Hello,
I am working on creating a mutant endothelial cell line having Neomycin knock-in present,
I have designed my homology arms of 1kb size having neomycin in between them, I would like to know if anyone has worked on stable transfection and have used any kind of mammalian vector as backbone for Donor DNA.
Previously i thought of using PuC based vector as i don't need expression of any protein but i'm not sure whether it will work or not because of its small size.
(my insert size is approx 900-1000bp and homology arms is of 1kb, my approach is to select cells using G418 post 48 hrs of transfection)
Please comment with any name or paper that you find relevant.
Thankyou for your suggestions.
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