Improved specificity of TALE-based genome editing using an expanded RVD repertoire. Miller, J. C.; Zhang, L.; Xia, D. F.; Campo, J. J.; Ankoudinova, I. V.; Guschin, D. Y.; Babiarz, J. E.; Meng, X.; Hinkley, S. J.; Lam, S. C.; Paschon, D. E.; Vincent, A. I.; Dulay, G. P.; Barlow, K. A.; Shivak, D. A.; Leung, E.; Kim, J. D.; Amora, R.; Urnov, F. D.; Gregory, P. D.; and Rebar, E. J. Nature Methods, 12(5):465–471, May, 2015.
Improved specificity of TALE-based genome editing using an expanded RVD repertoire [link]Paper  doi  abstract   bibtex   
Transcription activator–like effector (TALE) proteins have gained broad appeal as a platform for targeted DNA recognition, largely owing to their simple rules for design. These rules relate the base specified by a single TALE repeat to the identity of two key residues (the repeat variable diresidue, or RVD) and enable design for new sequence targets via modular shuffling of these units. A key limitation of these rules is that their simplicity precludes options for improving designs that are insufficiently active or specific. Here we address this limitation by developing an expanded set of RVDs and applying them to improve the performance of previously described TALEs. As an extreme example, total conversion of a TALE nuclease to new RVDs substantially reduced off-target cleavage in cellular studies. By providing new RVDs and design strategies, these studies establish options for developing improved TALEs for broader application across medicine and biotechnology.
@article{miller_improved_2015,
	title = {Improved specificity of {TALE}-based genome editing using an expanded {RVD} repertoire},
	volume = {12},
	copyright = {© 2015 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.},
	issn = {1548-7091},
	url = {http://www.nature.com/nmeth/journal/v12/n5/full/nmeth.3330.html},
	doi = {10.1038/nmeth.3330},
	abstract = {Transcription activator–like effector (TALE) proteins have gained broad appeal as a platform for targeted DNA recognition, largely owing to their simple rules for design. These rules relate the base specified by a single TALE repeat to the identity of two key residues (the repeat variable diresidue, or RVD) and enable design for new sequence targets via modular shuffling of these units. A key limitation of these rules is that their simplicity precludes options for improving designs that are insufficiently active or specific. Here we address this limitation by developing an expanded set of RVDs and applying them to improve the performance of previously described TALEs. As an extreme example, total conversion of a TALE nuclease to new RVDs substantially reduced off-target cleavage in cellular studies. By providing new RVDs and design strategies, these studies establish options for developing improved TALEs for broader application across medicine and biotechnology.},
	language = {en},
	number = {5},
	urldate = {2016-04-06},
	journal = {Nature Methods},
	author = {Miller, Jeffrey C. and Zhang, Lei and Xia, Danny F. and Campo, John J. and Ankoudinova, Irina V. and Guschin, Dmitry Y. and Babiarz, Joshua E. and Meng, Xiangdong and Hinkley, Sarah J. and Lam, Stephen C. and Paschon, David E. and Vincent, Anna I. and Dulay, Gladys P. and Barlow, Kyle A. and Shivak, David A. and Leung, Elo and Kim, Jinwon D. and Amora, Rainier and Urnov, Fyodor D. and Gregory, Philip D. and Rebar, Edward J.},
	month = may,
	year = {2015},
	keywords = {Biotechnology, Genetic engineering},
	pages = {465--471}
}
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