MG

@betowbin @FluoProt @joachimgoedhart Not a single point mutation but the spaghetti monster FPs mutated the 65-67 SYG of GFP to GGG to create dark variants nature.com/articles/nmeth…

Fluorescence-friends: I want to introduce a single point mutation in GFP that kills its fluorescence, but keeps its structure. What should I do? Where should I look? @FluoProt @joachimgoedhart

@SamantLab @AndrewWoodLab It’s pretty tough to have a stable C-terminal tag for Ub though

@AndrewWoodLab Depends what you mean by “don’t tolerate”—I’d argue that Ubiquitin shouldn’t be N- or C- terminal tagged because poly-Ub chains should be able to form from both termini (with the discovery of M1-Ub). But if you are asking for proteins that aren’t stable with tags, then no, sorry!

A Twitter hive mind question for protein engineers: I’m looking for examples of proteins that don’t tolerate tags (epitope, FP, degron etc.) at N- or C-termini. If you know of good examples, let’s hear them! RT’s appreciated.

@PShannonNeuro Hamartoma didn't fit with the aneuploidy anyways. A. Olives, prominent because of lack of pyramids. But are really prominent. B. Histology? C. Dorsal cyst wall from a better dissector than me. D. Fused thalami E. Choroid plexus Any polydactyly?

@PShannonNeuro Let’s see if I’m on the right track. B. Hypothalamic hamartoma. D. Fused/single superior colliculus

@PShannonNeuro @gemistocyte I think a genetic cause is odd for left-sided unilateral PMG. Isolated left MCA ischaemia? Maternal ischemia or infectious would be bilateral. Is there a vascular lesion/abnormality?

@PShannonNeuro Hemimegalencephaly. Look for cutaneous vascular lesions, poly/syndactyly.