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. Author manuscript; available in PMC: 2017 Nov 30.
Published in final edited form as: J Genet Genomics. 2016 Apr 24;43(5):239–250. doi: 10.1016/j.jgg.2016.04.008

Figure 1.

Figure 1

Design of synthetic sgRNA for S. pyogenes and S. aureus Cas9

A: The natural S. pyogenes guide RNA composed of crRNA and tracrRNA. B–D: Original design of Jinek et al. (2012) (B), standard, commonly used (C) and optimized (D) designs of S. pyogenes sgRNA. E: The natural S. aureus guide RNA. F–I: Original (F–H) and optimized (I) designs of S. aureus sgRNA. Red indicates the different sequences introduced to connect crRNA and tracrRNA in synthetic S. pyogenes and S. Aureus sgRNA. Mutations in the TTTT stretch of crRNA and the corresponding AAAA stretch of tracrRNA, which were made to improve sgRNA expressim from U6 promoter, are also indicated in red. sgRNA design impacts cleavage efficiency as indicated.