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Mining Endonuclease Cleavage Determinants in Genomic Sequence Data
Homing endonucleases have great potential as tools for targeted gene therapy and gene correction, but identifying variants of these enzymes capable of cleaving specific DNA targets of interest is necessary before the widespread use of such technologies is possible. We identified homologues of the LA...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3173205/ https://www.ncbi.nlm.nih.gov/pubmed/21778233 http://dx.doi.org/10.1074/jbc.M111.259572 |
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author | Szeto, Mindy D. Boissel, Sandrine J. S. Baker, David Thyme, Summer B. |
author_facet | Szeto, Mindy D. Boissel, Sandrine J. S. Baker, David Thyme, Summer B. |
author_sort | Szeto, Mindy D. |
collection | PubMed |
description | Homing endonucleases have great potential as tools for targeted gene therapy and gene correction, but identifying variants of these enzymes capable of cleaving specific DNA targets of interest is necessary before the widespread use of such technologies is possible. We identified homologues of the LAGLIDADG homing endonuclease I-AniI and their putative target insertion sites by BLAST searches followed by examination of the sequences of the flanking genomic regions. Amino acid substitutions in these homologues that were located close to the target site DNA, and thus potentially conferring differences in target specificity, were grafted onto the I-AniI scaffold. Many of these grafts exhibited novel and unexpected specificities. These findings show that the information present in genomic data can be exploited for endonuclease specificity redesign. |
format | Online Article Text |
id | pubmed-3173205 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-31732052011-09-21 Mining Endonuclease Cleavage Determinants in Genomic Sequence Data Szeto, Mindy D. Boissel, Sandrine J. S. Baker, David Thyme, Summer B. J Biol Chem Enzymology Homing endonucleases have great potential as tools for targeted gene therapy and gene correction, but identifying variants of these enzymes capable of cleaving specific DNA targets of interest is necessary before the widespread use of such technologies is possible. We identified homologues of the LAGLIDADG homing endonuclease I-AniI and their putative target insertion sites by BLAST searches followed by examination of the sequences of the flanking genomic regions. Amino acid substitutions in these homologues that were located close to the target site DNA, and thus potentially conferring differences in target specificity, were grafted onto the I-AniI scaffold. Many of these grafts exhibited novel and unexpected specificities. These findings show that the information present in genomic data can be exploited for endonuclease specificity redesign. American Society for Biochemistry and Molecular Biology 2011-09-16 2011-07-21 /pmc/articles/PMC3173205/ /pubmed/21778233 http://dx.doi.org/10.1074/jbc.M111.259572 Text en © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) applies to Author Choice Articles |
spellingShingle | Enzymology Szeto, Mindy D. Boissel, Sandrine J. S. Baker, David Thyme, Summer B. Mining Endonuclease Cleavage Determinants in Genomic Sequence Data |
title | Mining Endonuclease Cleavage Determinants in Genomic Sequence Data |
title_full | Mining Endonuclease Cleavage Determinants in Genomic Sequence Data |
title_fullStr | Mining Endonuclease Cleavage Determinants in Genomic Sequence Data |
title_full_unstemmed | Mining Endonuclease Cleavage Determinants in Genomic Sequence Data |
title_short | Mining Endonuclease Cleavage Determinants in Genomic Sequence Data |
title_sort | mining endonuclease cleavage determinants in genomic sequence data |
topic | Enzymology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3173205/ https://www.ncbi.nlm.nih.gov/pubmed/21778233 http://dx.doi.org/10.1074/jbc.M111.259572 |
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