title: Homing endonuclease target site specificity defined by sequential enrichment and next-generation sequencing of highly complex target site libraries.
abstract: Homing endonucleases (HEs) are DNA sequence-specific enzymes that recognize and cleave long target sites (14-40 bp) to generate double-strand breaks (DSBs). Their high site recognition specificity and tight coupling of binding and cleavage make HEs attractive reagents for targeted genome manipulation. In order to delineate the target site specificity of HEs and facilitate HE engineering, we have developed a method for comprehensive target site profiling of HEs cleavage specificity using partially randomized target site libraries and high-throughput DNA sequencing.
authors: Department of Pathology, University of Washington, Seattle, WA, USA.
authors: Hui
authors: H
authors: Li
authors: Raymond J
authors: RJ
authors: Monnat
mesh_terms: D001665:Binding Sites; D053837:DNA Cleavage; D004720:Endonucleases / Q000378:metabolism*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing; D013379:Substrate Specificity
chemical_list: D004720:Endonucleases