Team:Waterloo
Simple sgRNA Exchange
Cas9 PAM Flexibility
CRISPR Plant Defense
Re-engineering CRISPR-Cas9 with functional applications in eukaryotic systems
CRISPR-Cas9 is an exciting tool for synthetic biologists because it can target and edit genomes with unprecedented specificity. Our team is attempting to re-engineer CRISPR to make it more flexible and easier to use.
Simple sgRNA Exchange: We’re making it easy to test different sgRNA designs: restriction sites added to the sgRNA backbone allow 20 nucleotide target sequences to be swapped without excessive cloning. We aim to reduce the turnaround time for using CRISPR to target different sequences.
Cas9 PAM Flexibility: We’re applying recent research on viable mutations within Cas9’s PAM-interacting domain to design (d)Cas9 variants that bind to novel PAM sites, moving towards the goal of a suite of variants that can bind any desired sequence. We believe our re-engineered CRISPR-Cas9 will give biologists increased ability to optimize targeting in many applications.
CRISPR Plant Defense: The application we chose to explore is a proof-of-concept antiviral system defending the model plant Arabidopsis thaliana against Cauliflower Mosaic Virus, which would benefit from testing a large number of possible sgRNAs in the viral genome.
Jamboree Achievements
Our team had a fantastic time at the Jamboree this year, and we were more successful this year than ever before!
- Gold Medal Standing for the third year in a row
- Best Overgrad Software Tool
- Best Overgrad Poster (view here)
- Nominated for Best Foundational Advance Project