A transcription activator-like effector toolbox for genome engineering.
Transcription activator-like effectors (TALEs) are a class of naturally occurring DNA-binding proteins found in the plant pathogen Xanthomonas sp. The DNA-binding domain of each TALE consists of tandem 34-amino acid repeat modules that can be rearranged according to a simple cipher to target new DNA sequences. Customized TALEs can be used for a wide variety of genome engineering applications, including transcriptional modulation and genome editing. Here we describe a toolbox for rapid construction of custom TALE transcription factors (TALE-TFs) and nucleases (TALENs) using a hierarchical ligation procedure. This toolbox facilitates affordable and rapid construction of custom TALE-TFs and TALENs within 1 week and can be easily scaled up to construct TALEs for multiple targets in parallel. We also provide details for testing the activity in mammalian cells of custom TALE-TFs and TALENs using quantitative reverse-transcription PCR and Surveyor nuclease, respectively. The TALE toolbox described here will enable a broad range of biological applications.
Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, USA.
Neville E
NE
Sanjana
Le
L
Cong
Yang
Y
Zhou
Margaret M
MM
Cunniff
Guoping
G
Feng
Feng
F
Zhang
D000818:Animals; D005786:Gene Expression Regulation; D005818:Genetic Engineering / Q000379:methods*; D023281:Genomics / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D020080:Tandem Repeat Sequences; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D014974:Xanthomonas / Q000235:genetics*
D014157:Transcription Factors