Team:Waterloo/Lab/sgRNA
sgRNA Modification
sgRNA is a chimera of tracrRNA and crRNA, which were the original two-part biomolecule that allowed Cas9 to find and identify its target dsDNA. The purpose of fusing together the tracrRNA and crRNA was to simplify the CRISPR-Cas9 system for scientists to use more easily. Aligning with what the University of Waterloo is best known for, the lab team continued this innovation by spending part of its summer re-engineering sgRNA so that the twenty base pair target region could be removed and replaced by a new twenty base pair target region with basic classical cloning techniques. This challenge was tackled by first investigating the architecture of the sgRNA scaffold region, followed by a study of how different point mutations to the sgRNA scaffold region would affect its secondary structure. Both tasks were completed by a thorough scientific literature review that lead to the final, characterized design shown further below.