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The Generation of Zinc Finger Proteins by Modular Assembly

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Engineered Zinc Finger Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 649))

Abstract

The modular assembly (MA) method of generating engineered zinc finger proteins (ZFPs) was the first practical method for creating custom DNA-binding proteins. As such, MA has enabled a vast exploration of sequence-specific methods and reagents, ushering in the modern era of zinc finger-based applications that are described in this volume. The first zinc finger nuclease to cleave an endogenous site was created using MA, as was the first artificial transcription factor to enter phase II clinical trials. In recent years, other excellent methods have been developed that improved the affinity and specificity of the engineered ZFPs. However, MA is still used widely for many applications. This chapter will describe methods and give guidance for the creation of ZFPs using MA. Such ZFPs might be useful as starting materials to perform other methods described in this volume. Here, we also describe a single-strand annealing recombination assay for the initial testing of zinc finger nucleases.

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Acknowledgments

This chapter is based upon work supported by grant CA103651 from the National Cancer Institute, NIH (DJS).

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Authors and Affiliations

  1. Department of Pharmacology, Genome Center, University of California at Davis, Davis, CA, USA

    Mital S. Bhakta & David J. Segal

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  1. Mital S. Bhakta
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  2. David J. Segal
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Correspondence to David J. Segal .

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Editors and Affiliations

  1. School of Molecular &, Microbial Biosciences, University of Sydney, Cnr. Butlin Ave. & Maze Cres., Sydney, 2006, New South Wales, Australia

    Joel P. Mackay

  2. Dept. Pharmacology & Genome Center, University of California, Davis, E. Health Sciences Drive 451, Davis, 95616, California, USA

    David J. Segal

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Bhakta, M.S., Segal, D.J. (2010). The Generation of Zinc Finger Proteins by Modular Assembly. In: Mackay, J., Segal, D. (eds) Engineered Zinc Finger Proteins. Methods in Molecular Biology, vol 649. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-753-2_1

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  • DOI: https://doi.org/10.1007/978-1-60761-753-2_1

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-752-5

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