PIGN-CDG families are funding yeast-powered drug repurposing screens and a fibroblast biobank for hit validation. We just completed a pilot repurposing screen, and an expanded screen is in progress.
Thanks for the article. It is good to see updates of all the hard work you are doing. I appreciate your science and patience.
I have a clarifying question, I hope that you will see this and answer it for me.
My question is about your pivot to the temperature-sensitive (ts) yeast mutants. I am a little confused by your description here. Did you engineer-in the 10 PIGN variants described above into the ts yeast, and then run those Pharmakon screens? Or is your data comparing ts yeast to non-ts yeast? Or is it something else that I am missing?
Thanks for the help and reply. I am looking forward to hearing more about this work.
We transformed a MCD4 DaMP strain that has ~10% residual expression with plasmids containing the 10 PIGN variants. The MCD4 DaMP strain is slow growing.
The temperature-sensitive yeast mutant is totally separate. We did not express the 10 PIGN variants in the ts mutant. Like you, we considered the idea but for cost and time considerations we did not pursue it.
The ts mutant is compared to a wildtype yeast strain.
Thanks for the quick and helpful response, you have answered my question :)
I enjoy seeing some of the data, and hearing about both the progress and roadbumps along the way. Major props for keeping the conversation going in the comments, too. Good luck with the planned screens and continued work.
Thanks for the article. It is good to see updates of all the hard work you are doing. I appreciate your science and patience.
I have a clarifying question, I hope that you will see this and answer it for me.
My question is about your pivot to the temperature-sensitive (ts) yeast mutants. I am a little confused by your description here. Did you engineer-in the 10 PIGN variants described above into the ts yeast, and then run those Pharmakon screens? Or is your data comparing ts yeast to non-ts yeast? Or is it something else that I am missing?
Thanks for the help and reply. I am looking forward to hearing more about this work.
Thanks for your question! Happy to clarify below.
We transformed a MCD4 DaMP strain that has ~10% residual expression with plasmids containing the 10 PIGN variants. The MCD4 DaMP strain is slow growing.
The temperature-sensitive yeast mutant is totally separate. We did not express the 10 PIGN variants in the ts mutant. Like you, we considered the idea but for cost and time considerations we did not pursue it.
The ts mutant is compared to a wildtype yeast strain.
Thanks for the quick and helpful response, you have answered my question :)
I enjoy seeing some of the data, and hearing about both the progress and roadbumps along the way. Major props for keeping the conversation going in the comments, too. Good luck with the planned screens and continued work.