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Review
. 2017 Nov 29;18(12):2565.
doi: 10.3390/ijms18122565.

Applications of Alternative Nucleases in the Age of CRISPR/Cas9

Tuhin K Guha  1 , David R Edgell  2
Affiliations
Review

Applications of Alternative Nucleases in the Age of CRISPR/Cas9

Tuhin K Guha et al. Int J Mol Sci. .

Abstract

Breakthroughs in the development of programmable site-specific nucleases, including zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), meganucleases (MNs), and most recently, the clustered regularly interspaced short palindromic repeats (CRISPR) associated proteins (including Cas9) have greatly enabled and accelerated genome editing. By targeting double-strand breaks to user-defined locations, the rates of DNA repair events are greatly enhanced relative to un-catalyzed events at the same sites. However, the underlying biology of each genome-editing nuclease influences the targeting potential, the spectrum of off-target cleavages, the ease-of-use, and the types of recombination events at targeted double-strand breaks. No single genome-editing nuclease is optimized for all possible applications. Here, we focus on the diversity of nuclease domains available for genome editing, highlighting biochemical properties and the potential applications that are best suited to each domain.

Keywords: CRISPR/Cas9; FokI; GIY-YIG nuclease domain; TALEN; ZFN; dimeric nuclease; monomeric nuclease.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
DNA double-strand break repair pathways. (A) Homology dependent repair, applicable in the presence of a homologous repair template (green) and error-free non-homologous end joining pathway. The later, however, is responsible for the regeneration of the target site through religation of the paired end complex. Red lightning symbol indicates introduction of a DSB. Large red arrow highlights efficient repair and regeneration of the nuclease target site by NHEJ. (B) Schematic model representing how TevCas9 can bias DNA repair outcome (see text for details). Red dashed rectangles, endonuclease target site; green dashed rectangles, mutated target site after error-prone repair. NHEJ, non-homologous end joining; HDR, homology directed repair; cNHEJ, classical NHEJ; alt-NHEJ, alternative-NHEJ.
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