Figure 1. A schematic presentation of a ribozyme-flanked gRNA (RGR, ribozyme-gRNA-ribozyme) molecule.

Once transcribed, the corresponding RGR DNA will produce an RNA molecule with a ribozyme at both the 5′- and 3′-end. The primary transcripts will undergo self-cleavage to release the mature gRNA. The RGR design allows the production of a functional gRNA molecule from any promoter, enabling production of gRNA molecules in vitro, in vivo, and in tissue/cell specific manner.