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.
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Acknowledgments
This work was supported by US National Institutes of Health Training Grant award to H.L. (5RL9HL092555); by a US National Institutes of Health U54 Interdisciplinary Research Roadmap award (1RL1 CA133831) to R.J.M. Jr; and by a grant from the Foundation for the National Institutes of Health funded by the Bill and Melinda Gates Foundation as a Grand Challenges in Global Health award to R.J.M. Jr.
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Li, H., Monnat, R.J. (2014). Homing Endonuclease Target Site Specificity Defined by Sequential Enrichment and Next-Generation Sequencing of Highly Complex Target Site Libraries. In: Edgell, D. (eds) Homing Endonucleases. Methods in Molecular Biology, vol 1123. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-968-0_12
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DOI: https://doi.org/10.1007/978-1-62703-968-0_12
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