Article open access publication

Engineering a Nickase on the Homing Endonuclease I-DmoI Scaffold

Journal of Biological Chemistry, American Society for Biochemistry & Molecular Biology (ASBMB), ISSN 0021-9258

Volume 290, 30, 2015

DOI:10.1074/jbc.m115.658666, Dimensions: pub.1047103685, PMC: PMC4513113, PMID: 26045557,

Affiliations

Organisations

  1. (1) From the ‡Macromolecular Crystallography Group and
  2. (2) Spanish National Cancer Research Centre, grid.7719.8
  3. (3) Takara (France), grid.433267.7
  4. (4) Sapienza University of Rome, grid.7841.a
  5. (5) University of Copenhagen, grid.5254.6, KU

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Italy

France

Denmark

Spain

Continents

Europe

Description

Homing endonucleases are useful tools for genome modification because of their capability to recognize and cleave specifically large DNA targets. These endonucleases generate a DNA double strand break that can be repaired by the DNA damage response machinery. The break can be repaired by homologous recombination, an error-free mechanism, or by non-homologous end joining, a process susceptible to introducing errors in the repaired sequence. The type of DNA cleavage might alter the balance between these two alternatives. The use of "nickases" producing a specific single strand break instead of a double strand break could be an approach to reduce the toxicity associated with non-homologous end joining by promoting the use of homologous recombination to repair the cleavage of a single DNA break. Taking advantage of the sequential DNA cleavage mechanism of I-DmoI LAGLIDADG homing endonuclease, we have developed a new variant that is able to cut preferentially the coding DNA strand, generating a nicked DNA target. Our structural and biochemical analysis shows that by decoupling the action of the catalytic residues acting on each strand we can inhibit one of them while keeping the other functional.

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University of Copenhagen

Dimensions Citation Indicators

Times Cited: 7

Field Citation Ratio (FCR): 0.94

Relative Citation ratio (RCR): 0.36

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