Hi guys,

I am going to knock-in a donor template (HA-left // transgene cassette // HA-right) into a CRISPR/Cas9-mediated DSB.

Homology arm (HA) left = 3.5kb

Transgene cassette = 6.5kb

Homology arm (HA) right = 2.5kb

The homology arm right has 1 mutation towards the outside (away from the transgene), which does not mess with anything important in the genome. However, I'm now wondering whether this one mutation has any chance to interfere with my knock-in?

My guess would be that it won't matter, or at most will reduce knock-in efficiency a bit. Please let me know if you'd think otherwise and why. :-)

Kind regards