A transcription activator-like effector toolbox for genome engineering View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2012-01

AUTHORS

Neville E Sanjana, Le Cong, Yang Zhou, Margaret M Cunniff, Guoping Feng, Feng Zhang

ABSTRACT

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. More... »

PAGES

171

References to SciGraph publications

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  • Journal

    TITLE

    Nature Protocols

    ISSUE

    1

    VOLUME

    7

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nprot.2011.431

    DOI

    http://dx.doi.org/10.1038/nprot.2011.431

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1020679703

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/22222791


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