Skip Navigation



Nucleic Acids Research Advance Access published online on January 19, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp004
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (7326K) Freely available
Right arrow Screen PDF (939K) Freely available
Right arrow Supplementary Data
Right arrowOA All Versions of this Article:
37/5/1650    most recent
gkp004v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Li, H.
Right arrow Articles by Monnat, R. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Li, H.
Right arrow Articles by Monnat, R. J., Jr
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Nucleic Acid Enzymes

Generation of single-chain LAGLIDADG homing endonucleases from native homodimeric precursor proteins

Hui Li1,2, Stefan Pellenz1,2, Umut Ulge2,3, Barry L. Stoddard2,4 and Raymond J. Monnat, Jr*

1Department of Pathology, University of Washington, Box 357705, Seattle, WA 98195, 2Northwest Genome Engineering Consortium, Seattle, WA, 3Department of the Molecular and Cellular Biology Program, University of Washington, Box 357705, Seattle, WA 98195, 4Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue N. A3-025, Seattle, WA 98109 and 5Department of Genome Sciences, University of Washington, Box 357705, Seattle, WA 98195, USA

*To whom correspondence should be addressed. Tel: +1 206 616 7392; Fax: +1 206 543 3967; Email: monnat{at}u.washington.edu

Received November 26, 2008. Revised December 30, 2008. Accepted January 2, 2009.

Homing endonucleases (HEs) cut long DNA target sites with high specificity to initiate and target the lateral transfer of mobile introns or inteins. This high site specificity of HEs makes them attractive reagents for gene targeting to promote DNA modification or repair. We have generated several hundred catalytically active, monomerized versions of the well-characterized homodimeric I-CreI and I-MsoI LAGLIDADG family homing endonuclease (LHE) proteins. Representative monomerized I-CreI and I-MsoI proteins (collectively termed mCreIs or mMsoIs) were characterized in detail by using a combination of biochemical, biophysical and structural approaches. We also demonstrated that both mCreI and mMsoI proteins can promote cleavage-dependent recombination in human cells. The use of single chain LHEs should simplify gene modification and targeting by requiring the expression of a single small protein in cells, rather than the coordinate expression of two separate protein coding genes as is required when using engineered heterodimeric zinc finger or homing endonuclease proteins.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
S. Grizot, J. Smith, F. Daboussi, J. Prieto, P. Redondo, N. Merino, M. Villate, S. Thomas, L. Lemaire, G. Montoya, et al.
Efficient targeting of a SCID gene by an engineered single-chain homing endonuclease
Nucleic Acids Res., September 1, 2009; 37(16): 5405 - 5419.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.