Description

The CRISPR/Cas9 system is used to directly edit the DNA of an organism. It is bacteria’s immune system against bacteriophages. Bacteria incorporate fragments of the virus genome and use them as a defense mechanism. Once the bacteria encounter the same virus, a guide RNA locates a specific section of the DNA and signals for the Cas9 protein to lock onto the DNA. The Cas9 then breaks both strands of DNA and can then be used to disable a gene or insert a new gene. This system allows for easier and faster modification of the genome than current methods; this technology can help scientists and the public in many ways, such as finding cures for genetic therapy, understanding the role of proteins within cell, cell function, and genetic diseases.Our goal is to design and characterize an assay that allows scientists to measure the binding efficiencies of separate CRISPR Cas9 systems. Our model relies on competition between two Cas9s from two separate bacterial species. In each experiment, one Cas9 will be active, while the other will deactivated. Binding of the active Cas9 will cleave the testing plasmid, which instructs the cell to make Green Fluorescent Protein. Better binding of the active Cas9 will thus lead to degradation of the plasmid, making cells less green. Better binding by the deactivated Cas9 won’t affect the plasmid, but it will prevent the active Cas9 from binding and causing the plasmid to degrade, so the cells will stay green.