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. Author manuscript; available in PMC: 2018 Oct 30.
Published in final edited form as: Cell Stem Cell. 2017 Oct 5;21(4):431–447. doi: 10.1016/j.stem.2017.09.006

Figure 2. Technical considerations to improve CRISPR/Cas9 versatility.

Figure 2.

Several factors can improve the efficiency of the epigenetic targeted modifications by CRISPR/Cas9. A) Using only the effector domain of the chromatin-modifiers, as well as binding peptidic linkers and nuclear localization signals (NLS) to the dCas9 enzyme, significantly increases the precision and efficiency of the intended chromatin modifications. B) The CRISPR/Cas9 system offers 4 different options to amplify the effect of single chromatin modifications, based on the possibility to multiplex the amount of effectors positioned in a specific locus. C) The CRISPR/Cas9 system offers the possibility to simultaneously induce several targeted chromatin modifications, as the gRNA molecules can function as scaffolds to recruit different RNA-binding proteins linked to various effector domains. Another option to target multiple loci is to exploit the orthogonality of this system, using Cas9 enzymes from different species and their corresponding gRNAs. Sp (Streptococcus pyogenes), St (Streptococcus thermophiles), Nm (Neisseria meningitides), Sa (Staphylococcus aureus).