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Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications.


ABSTRACT: 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.

SUBMITTER: Li H 

PROVIDER: S-EPMC3315327 | biostudies-literature | 2012 Mar

REPOSITORIES: biostudies-literature

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Comprehensive homing endonuclease target site specificity profiling reveals evolutionary constraints and enables genome engineering applications.

Li Hui H   Ulge Umut Y UY   Hovde Blake T BT   Doyle Lindsey A LA   Monnat Raymond J RJ  

Nucleic acids research 20111125 6


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  ...[more]

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