Team:Waterloo
OFF to priON
Using stop codon read-through and CRISPR to explore S. cerevisiae prion mechanisms
Furthering the advancement of research into prions and neurodegenerative diseases
We initiate a [PSI+] response in S. cerevisiae using a plasmid that will overexpress Sup35-NM.
We insert a premature stop codon into an N-terminal CFP tag that will be read through and fluoresce during a [PSI+] response
We control read through rates by choosing the location of the premature stop codon, and can therefore control protein production in [PSI+]. We use Hsp104, a chaperone protein in S. cerevisiae, to demonstrate that our set-up phenotypically responds to the stop codon readthrough.
