Split mCherry

Posted on by kmatreyek

I had a product that could have benefitted from using split mCherry to serve an AND function. Put the split mCherry in my usual mCherry spot in the recombination vector (2A’d with Puromycin, also), and I couldn’t see any visible fluorescence when both the small and big fragments were in the same cell. After some time, I saw a paper using split super-folder mCherry fused with the Spycatcher system, so I used that and that seemed to allow us to now see a shift in fluorescence off the background. I found another paper using an improved split super-folder mCherry (sfmCherry3C), and tested that, which seemed to work slightly better.

So while not perfect, in that there isn’t *complete* separation from the background distribution, it’s shifted away enough that it should serve my purposes (for now). Though, well, I’ll probably keep playing around with this (perhaps adding something like a leucine zipper?) and seeing if that helps increase fluorescence.