Does anyone know of nickases that are non-specific and are functional as monomers? Something like FokI, but FokI can only cut as a dimer. I know of an instance where cleavase TevI was fused at C terminus of a TALE such that it turned into a nickase (https://www.nature.com/articles/ncomms2782), something like would also work, as I want to fuse it to another protein. But TevI is semi-specific. Thank you
Hi..
Yes, there are several examples of non-specific nickases that function as monomers. One example is the enzyme Cas9, which is a member of the CRISPR-Cas family of nucleases. Cas9 is a non-specific endonuclease that can be programmed to recognize specific DNA sequences and introduce double-stranded breaks (DSBs) at those locations. Unlike FokI, which requires dimerization to function, Cas9 is active as a monomer and can introduce DSBs in the DNA even when it is not dimerized.
Another example of a non-specific nickase that functions as a monomer is the enzyme Cpf1, which is also a member of the CRISPR-Cas family. Like Cas9, Cpf1 can be programmed to recognize specific DNA sequences and introduce DSBs at those locations, but it has a slightly different mechanism of action and can be more efficient at generating precise cuts in certain contexts.
I hope this information is helpful!
Regards
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