Figure 3.

Overview and comparison of the utilisation of circularised guide RNAs. Both circular gRNA are generated with the help of autocatalytically active twister ribozymes and vary in the length of their antisense domain. To improve target efficiency, the antisense region can be lengthened, or the “dose” of the gRNA increased, or the structure of the gRNA made more flexible by including a linker sequence. To decrease bystander editing, deletions, loops and A-G mismatches are used to disrupt perfectly paired gRNA. Circular gRNA can be delivered as DNA or RNA.