Is it any better enzyme to use? Is there different type of this enzyme that we can use?
Hi there,
The main point in using FokI endonuclease domain is that it is not DNA sequence specific (DNA recognition being defined by TALE domain) and that it acts as a dimer to ensure DNA ds break. Any other endonuclease bearing the same properties may also be used in theory.
No single genome-editing nuclease can work optimally for all applications. 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. Therefore, other gene editing nucleases that could be involved in gene editing in place of FOK1 could include monomeric nuclease, dimeric nuclease, GIY-YIG nuclease domain and CRISPR/Cas9
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