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TAL effector RVD specificities and efficiencies

Nature Biotechnology volume 30, pages 593–595 (2012)Cite this article

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To the Editor:

The DNA-binding domain of transcription activator–like effectors (TALEs) has become an important tool for the programmable and specific targeting of genome-editing nucleases1,2,3,4. We determined specificities of TALE DNA-binding modules and discovered varying efficiencies to promote TALE activity. This prompted us to study the basis of TALE efficiency. Natural TALEs from plant-pathogenic Xanthomonas sp. bacteria function as eukaryotic transcription factors. TALEs bind to DNA via tandem near-identical 34-amino-acid repeats, each of which recognizes one base pair via two adjacent amino acids (12 and 13) termed repeat-variable diresidues (RVDs)5,6. Three-dimensional structures of TALE–DNA complexes revealed that only amino acid 13 mediates specific recognition of the sense-strand DNA base, whereas amino acid 12 stabilizes the repeat structure7,8. In vitro assembly of repeat units yielded TALEs with new and predictable DNA specificities2,9. Different DNA-recognition specificities of individual RVDs have been shown4,5,10. Specific recognition of guanine is difficult because the common RVD NN (Asn-Asn) recognizes guanine and adenine, whereas the guanine-specific RVD NK (Asn-Lys) apparently functions less well than NN does11. Here we describe new RVD specificities including NH (Asn-His), which is highly specific for guanine. We show that efficiency of RVDs varies and that strong repeats (HD (His-Asn) or NN) are key for overall activity of TALEs.

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Figure 1: TALE RVD specificities and efficiencies.

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Acknowledgements

We thank R. Szczesny and T. Schreiber for helpful discussions on the manuscript, S. Thieme (Department of Genetics, Martin Luther University Halle-Wittenberg) for providing the binary vector, C. Kretschmer for expert technical assistance and U. Bonas for continued support. J.B. was supported by a grant from the Deutsche Forschungsgemeinschaft (SPP 1212).

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  1. Department of Genetics, Martin Luther University Halle-Wittenberg, Halle (Saale), Germany

    Jana Streubel, Christina Blücher, Angelika Landgraf & Jens Boch

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  1. Jana Streubel
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  2. Christina Blücher
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  3. Angelika Landgraf
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  4. Jens Boch
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Correspondence to Jens Boch.

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J.B. is a part owner of a patent application regarding the use of TALs.

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Streubel, J., Blücher, C., Landgraf, A. et al. TAL effector RVD specificities and efficiencies. Nat Biotechnol 30, 593–595 (2012). https://doi.org/10.1038/nbt.2304

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