The CRISPR/Cas9 system is widely used for various genome editing approaches in cultured cells and living organisms and was broadly explored for preclinical applications. This two component system is composed of the Cas9 endonuclease that acts in cooperation with a chimeric guide RNA (gRNA) mediating the sequence-specific binding to its complementary target protospacer sequence preceding a protospacer adjunct motif (PAM). Because of its simple gRNA design and easy cloning procedure for customization, the CRISPR/Cas9 system is easier to handle than transcription activator-like effector nucleases (TALENs) and artificial zinc finger nucleases (ZFN). As veritable gene delivery vehicles, viral vectors could be particularly fit to broaden the applicability of RGNs to other cell types including dividing and quiescent primary cells.

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