Programmable nucleases — including zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and RNA-guided engineered nucleases (RGENs) derived from the bacterial clustered regularly interspaced short palindromic repeat (CRISPR)–Cas (CRISPR-associated) system — enable targeted genetic modifications in cultured cells, as well as in whole animals and plants. The value of these enzymes in research, medicine and biotechnology arises from their ability to induce site-specific DNA cleavage in the genome, the repair (through endogenous mechanisms) of which allows high-precision genome editing. However, these nucleases differ in several respects, including their composition, targetable sites, specificities and mutation signatures, among other characteristics. Knowledge of nuclease-specific features, as well as of their pros and cons, is essential for researchers to choose the most appropriate tool for a range of applications.
At a glance
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This study shows that ZFNs can be delivered in vivo and lead to gene correction in mice.
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This milestone paper in the field of genome editing presents efficient gene correction using ZFNs in human cell lines.
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References 75 and 76 present the code of TALE–DNA interactions and paved the way for the construction of TALENs.
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This paper presents a genome-scale library of TALENs that target human miRNA sequences.
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References 23, 36 and 109 present paired Cas9 nickases that produce two offset SSBs on opposite DNA strands to enhance the specificity of RNA-guided genome editing, which is a strategy originally implemented using paired ZFNickases (reference 142).
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This paper shows that the Cas9 protein is guided by small RNAs to cleave DNA in a targeted manner in vitro, which paved the way for RNA-guided genome editing both in cells and in vivo.
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