Since it’s binding based, it makes sense you can delete these regions but it is neat that you dont majorly affect positioning using the typical sgRNA length, though circular permutation might yield different results overall. Thanks for the detailed insights, and best of luck with your submission! 😄

Also its so interesting seeing how robust these proteins are to large scale deletions like this, as you would imagine the spacing of the domains themselves to one another would greatly impact your ability to truncate them. Did you guys also consider playing w/ sgRNA content like in the OG paper? 😄

Hahaha also although you can’t edit, you can at least delete posts here as I am sure people on my feed noticed as I was writing on my preprint with the same typo 5 times in a row 🤦

Thanks for the full response Andrew! I suppose it makes sense you’re mostly optimising for ease/time here although you still need to order the sgRNA libraries. As for small deletions, I certainly agree but I also imagine having connective residues could help string deleted junctions together

You know I liked this paper, but I was more curious why they went this route for library construction versus trying to improve their earlier MISER strategy given that recombineering efficiency has seen some advances since. Maybe someone @savagecatsonly.bsky.social can help me 😄

I’ll eventually get caught up, but I listen while pipetting so there’s a lot of “day science” to get through first (right now on recording 21 with Daniel Kahneman 😅😂)

Really interesting and impressive work!!!