Protein Engineering Strategies to Expand CRISPR-Cas9 Applications

Figure 2

Schematic representations of Cas9 engineering by domain fusion. (a) End-to-end gene fusion approach. A diverse range of new technologies for DNA manipulation can be created by end-to-end fusion of Cas9 with an active domain. These fusions have been used to expand Cas9 applications. (b) Schematic depiction of Cas9 engineering by domain insertion. A switch behaviour could emerge in such a way that the Cas9 protein could be regulated by the input domain’s recognition of an input signal. Either binding domains or inteins have been used to generate Cas9 with a switch response. DNA sequences are depicted as lines. A light gray color of the Cas9 indicates that the protein is inactive or less active while blue represents the active state. The signal that modulates the switch is showed as a black triangle.

(a)

(b)