Campus Units
Genetics, Development and Cell Biology
Document Type
Article
Publication Version
Published Version
Publication Date
8-2010
Journal or Book Title
Nucleic Acids Research
Volume
39
Issue
1
First Page
359
Last Page
372
DOI
10.1093/nar/gkq704
Abstract
DNA double-strand breaks enhance homologous recombination in cells and have been exploited for targeted genome editing through use of engineered endonucleases. Here we report the creation and initial characterization of a group of rare-cutting, site-specific DNA nucleases produced by fusion of the restriction enzyme FokI endonuclease domain (FN) with the high-specificity DNA-binding domains of AvrXa7 and PthXo1. AvrXa7 and PthXo1 are members of the transcription activator-like (TAL) effector family whose central repeat units dictate target DNA recognition and can be modularly constructed to create novel DNA specificity. The hybrid FN-AvrXa7, AvrXa7-FN and PthXo1-FN proteins retain both recognition specificity for their target DNA (a 26 bp sequence for AvrXa7 and 24 bp for PthXo1) and the double-stranded DNA cleaving activity of FokI and, thus, are called TAL nucleases (TALNs). With all three TALNs, DNA is cleaved adjacent to the TAL-binding site under optimal conditions in vitro . When expressed in yeast, the TALNs promote DNA homologous recombination of a LacZ gene containing paired AvrXa7 or asymmetric AvrXa7/PthXo1 target sequences. Our results demonstrate the feasibility of creating a tool box of novel TALNs with potential for targeted genome modification in organisms lacking facile mechanisms for targeted gene knockout and homologous recombination.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License
Copyright Owner
The Authors
Copyright Date
2011
Language
en
File Format
application/pdf
Recommended Citation
Li, Ting; Huang, Sheng; Jiang, Wen Zhi; Wright, David; Spalding, Martin H.; Weeks, Donald P.; and Yang, Bing, "TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain" (2010). Genetics, Development and Cell Biology Publications. 174.
https://lib.dr.iastate.edu/gdcb_las_pubs/174
Comments
This article is published as Li, Ting, Sheng Huang, Wen Zhi Jiang, David Wright, Martin H. Spalding, Donald P. Weeks, and Bing Yang. "TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain." Nucleic acids research 39, no. 1 (2010): 359-372. 10.1093/nar/gkq704. Posted with permission.