Applications of Alternative Nucleases in the Age of CRISPR/Cas9

• Main Authors: Tuhin K. Guha, David R. Edgell
• Format: Article
• Language: English
• Published: MDPI AG 2017-11-01
• Series: International Journal of Molecular Sciences
• Subjects: monomeric nuclease; dimeric nuclease; GIY-YIG nuclease domain; FokI; CRISPR/Cas9; ZFN; TALEN
• Online Access: https://www.mdpi.com/1422-0067/18/12/2565

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.