Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting
University of Minnesota · University of Electronic Science and Technology of China · +2 more institutions
Abstract
TALENs are important new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors of plant pathogenic Xanthomonas spp. to the FokI nuclease, TALENs bind and cleave DNA in pairs. Binding specificity is determined by customizable arrays of polymorphic amino acid repeats in the TAL effectors. We present a method and reagents for efficiently assembling TALEN constructs with custom repeat arrays. We also describe design guidelines based on naturally occurring TAL effectors and their binding sites. Using software that applies these guidelines, in nine genes from plants, animals and protists, we found candidate cleavage sites on average every 35 bp. Each of 15 sites selected from this set…
Citation impact
- FWCI
- 247.84
- Percentile
- 100%
- References
- 30
Authors
10- TČTomáš ČermákCorresponding
University of Minnesota, University of Electronic Science and Technology of China, Iowa State University, University of Minnesota Rochester
- EDErin Doyle
University of Electronic Science and Technology of China, Iowa State University, University of Minnesota Rochester
- MCMichelle Christian
University of Electronic Science and Technology of China, Iowa State University, University of Minnesota Rochester
- LWLi Wang
University of Electronic Science and Technology of China, Iowa State University, University of Minnesota Rochester
- YZYong Zhang
University of Electronic Science and Technology of China, Iowa State University, University of Minnesota Rochester
Topics & keywords
- Transcription activator-like effector nuclease
- Biology
- Effector
- Genome editing
- Xanthomonas
- Computational biology
- Genetics
- Genome engineering