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Proc Natl Acad Sci U S A. 1996 Feb 6; 93(3): 1156–1160.
doi: 10.1073/pnas.93.3.1156
PMCID: PMC40048
PMID: 8577732

Hybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain.

Y G Kim, J Cha, and S Chandrasegaran
Author information Copyright and License information PMC Disclaimer

Abstract

A long-term goal in the field of restriction-modification enzymes has been to generate restriction endonucleases with novel sequence specificities by mutating or engineering existing enzymes. This will avoid the increasingly arduous task of extensive screening of bacteria and other microorganisms for new enzymes. Here, we report the deliberate creation of novel site-specific endonucleases by linking two different zinc finger proteins to the cleavage domain of Fok I endonuclease. Both fusion proteins are active and under optimal conditions cleave DNA in a sequence-specific manner. Thus, the modular structure of Fok I endonuclease and the zinc finger motifs makes it possible to create "artificial" nucleases that will cut DNA near a predetermined site. This opens the way to generate many new enzymes with tailor-made sequence specificities desirable for various applications.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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