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Chimeric Restriction Enzymes: What Is Next?
-
S. Chandrasegaran
Published/Copyright:
June 1, 2005
Abstract
Chimeric restriction enzymes are a novel class of engineered nucleases in which the non-specific DNA cleavage domain of FokI (a type IIS restriction endonuclease) is fused to other DNA-binding motifs. The latter include the three common eukaryotic DNA-binding motifs, namely the helix-turn-helix motif, the zinc finger motif and the basic helix-loop-helix protein containing a leucine zipper motif. Such chimeric nucleases have been shown to make specific cuts in vitro very close to the expected recognition sequences. The most important chimeric nucleases are those based on zinc finger DNA-binding proteins because of their modular structure. Recently, one such chimeric nuclease, Zif-QQR-FN was shown to find and cleave its target in vivo. This was tested by microinjection of DNA substrates and the enzyme into frog oocytes (Carroll et al., 1999). The injected enzyme made site-specific double-strand breaks in the targets even after assembly of the DNA into chromatin. In addition, this cleavage activated the target molecules for efficient homologous recombination. Since the recognition specificity of zinc fingers can be manipulated experimentally, chimeric nucleases could be engineered so as to target a specific site within a genome. The availability of such engineered chimeric restriction enzymes should make it feasible to do genome engineering, also commonly referred to as gene therapy.
:
Published Online: 2005-06-01
Published in Print: 1999-07-01
Copyright © 1999 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- Paul Nurse Felix Hoppe-Seyler Lecturer 1999
- Cyclin Dependent Kinases and Regulation of the Fission Yeast Cell Cycle
- Paper of the Year 1998
- Autonomous Regulation in Mammalian Mitochondrial DNA Transcription
- Prospects for the Precise Engineering of Plant Genomes by Homologous Recombination
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- Viroids with Hammerhead Ribozymes: Some Unique Structural and Functional Aspects with Respect to Other Members of the Group
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