Team:Waterloo

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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.

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
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