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Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications
Homing endonucleases (HEs) promote the evolutionary persistence of selfish DNA elements by catalyzing element lateral transfer into new host organisms. The high site specificity of this lateral transfer reaction, termed homing, reflects both the length (14–40 bp) and the limited tolerance of target...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3315327/ https://www.ncbi.nlm.nih.gov/pubmed/22121229 http://dx.doi.org/10.1093/nar/gkr1072 |
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author | Li, Hui Ulge, Umut Y. Hovde, Blake T. Doyle, Lindsey A. Monnat, Raymond J. |
author_facet | Li, Hui Ulge, Umut Y. Hovde, Blake T. Doyle, Lindsey A. Monnat, Raymond J. |
author_sort | Li, Hui |
collection | PubMed |
description | Homing endonucleases (HEs) promote the evolutionary persistence of selfish DNA elements by catalyzing element lateral transfer into new host organisms. The high site specificity of this lateral transfer reaction, termed homing, reflects both the length (14–40 bp) and the limited tolerance of target or homing sites for base pair changes. In order to better understand molecular determinants of homing, we systematically determined the binding and cleavage properties of all single base pair variant target sites of the canonical LAGLIDADG homing endonucleases I-CreI and I-MsoI. These Chlorophyta algal HEs have very similar three-dimensional folds and recognize nearly identical 22 bp target sites, but use substantially different sets of DNA-protein contacts to mediate site-specific recognition and cleavage. The site specificity differences between I-CreI and I-MsoI suggest different evolutionary strategies for HE persistence. These differences also provide practical guidance in target site finding, and in the generation of HE variants with high site specificity and cleavage activity, to enable genome engineering applications. |
format | Online Article Text |
id | pubmed-3315327 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-33153272012-03-30 Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications Li, Hui Ulge, Umut Y. Hovde, Blake T. Doyle, Lindsey A. Monnat, Raymond J. Nucleic Acids Res Nucleic Acid Enzymes Homing endonucleases (HEs) promote the evolutionary persistence of selfish DNA elements by catalyzing element lateral transfer into new host organisms. The high site specificity of this lateral transfer reaction, termed homing, reflects both the length (14–40 bp) and the limited tolerance of target or homing sites for base pair changes. In order to better understand molecular determinants of homing, we systematically determined the binding and cleavage properties of all single base pair variant target sites of the canonical LAGLIDADG homing endonucleases I-CreI and I-MsoI. These Chlorophyta algal HEs have very similar three-dimensional folds and recognize nearly identical 22 bp target sites, but use substantially different sets of DNA-protein contacts to mediate site-specific recognition and cleavage. The site specificity differences between I-CreI and I-MsoI suggest different evolutionary strategies for HE persistence. These differences also provide practical guidance in target site finding, and in the generation of HE variants with high site specificity and cleavage activity, to enable genome engineering applications. Oxford University Press 2012-03 2011-11-25 /pmc/articles/PMC3315327/ /pubmed/22121229 http://dx.doi.org/10.1093/nar/gkr1072 Text en © The Author(s) 2011. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Nucleic Acid Enzymes Li, Hui Ulge, Umut Y. Hovde, Blake T. Doyle, Lindsey A. Monnat, Raymond J. Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications |
title | Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications |
title_full | Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications |
title_fullStr | Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications |
title_full_unstemmed | Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications |
title_short | Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications |
title_sort | comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3315327/ https://www.ncbi.nlm.nih.gov/pubmed/22121229 http://dx.doi.org/10.1093/nar/gkr1072 |
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