Figure 2 : CRISPR-Cas design for eye gene of interest.

Shows how to design CRISPR-Cas for gene targeting. Adapted from the Addgene website (https://www.addgene.org/CRISPR/guide/). The short guide RNA and Cas9 endonuclease can be on the same or two different plasmids. Cas9 applications: endonuclease, nickase, by inactivating Cas9 catalytic domain- Cas9 can be used as a fusion protein containing DNA binding domain of Cas9 + transcriptional activator or repressor, cytidine or thymidine deaminase, the fusion of histone modifiers for epigenetic regulation, the fusion of fluorescent proteins for imaging of genomic loci.

Kawalpreet K Aneja Journal of Eye and Ophthalmology  2020 7:1DOI : 10.7243/2055-2408-7-1