I am trying to do a CRISPR/Cas9 knock-in and am wondering about the design of the donor vector (with the HDR template).
I've been looking through many publications, but never actually find an explanation how the backbone is chosen. If the vector consists of an ORI and an antibiotic selection marker, can you just take any backbone, like pUC19 or so on, or does a knock-in require certain plasmid designs (apart from the insertion cassette, of course)?
Nina Reuven
I have used pBluescript as a backbone, due to the same considerations you mentioned. It has worked fine for me. I also just assumed that any backbone without a eukaryotic promoter (for use in eukaryotic cells, in my case- mammalian) should work.
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