Results
Towards the end of our project, the Feng Zhang group at MIT published the crystal structure of saCas9 in Cell2. The paper demonstrated the development of a conditionally dimerizable saCas9 system using the FKBP-FRB domain pairs and PYL-ABI domain pairs, which is what we wanted to accomplish.
One facet worth noting is that the Zhang group used the 3D structure of the saCas9 protein to inform their split site choices. [what were their choices? what was their criteria?]
Out of the three split sites that they chose, the one that worked best was almost identical (one amino acid away) to one of the split sites that we identified using our model. Furthermore, the other two split sites that they chose which were not as successful happened to be located in regions with secondary structural elements.
This important result gave us some experimental validation that our model could offer scientists insights into choosing promising viable split sites for proteins, without only relying on 3D structures of proteins.
