Transcription Activator-Like Effectors (TALEs) Hybrid Nucleases for Genome Engineering Application
Type
ThesisAuthors
Wibowo, AnjarAdvisors
Zhu, Jian-Kang
Committee members
Gehring, Christoph A
Xiong, Liming

Program
BioscienceKAUST Department
Biological and Environmental Science and Engineering (BESE) DivisionDate
2011-06-06Permanent link to this record
http://hdl.handle.net/10754/209404
Metadata
Show full item recordAbstract
Gene targeting is a powerful genome engineering tool that can be used for a variety of biotechnological applications. Genomic double-strand DNA breaks generated by engineered site-specific nucleases can stimulate gene targeting. Hybrid nucleases are composed of DNA binding module and DNA cleavage module. Zinc Finger Nucleases were used to generate double-strand DNA breaks but it suffers from failures and lack of reproducibility. The transcription activator–like effectors (TALEs) from plant pathogenic Xanthomonas contain a unique type of DNA-binding domain that bind specific DNA targets. The purpose of this study is to generate novel sequence specific nucleases by fusing a de novo engineered Hax3 TALE-based DNA binding domain to a FokI cleavage domain. Our data show that the de novo engineered TALE nuclease can bind to its target sequence and create double-strand DNA breaks in vitro. We also show that the de novo engineered TALE nuclease is capable of generating double-strand DNA breaks in its target sequence in vivo, when transiently expressed in Nicotiana benthamiana leaves. In conclusion, our data demonstrate that TALE-based hybrid nucleases can be tailored to bind a user-selected DNA sequence and generate site-specific genomic double-strand DNA breaks. TALE-based hybrid nucleases hold much promise as powerful molecular tools for gene targeting applications.Citation
Wibowo, A. (2011). Transcription Activator-Like Effectors (TALEs) Hybrid Nucleases for Genome Engineering Application. KAUST Research Repository. https://doi.org/10.25781/KAUST-T2D61ae974a485f413a2113503eed53cd6c53
10.25781/KAUST-T2D61