The generation of zinc finger proteins by modular assembly

Mital S. Bhakta, David Segal

Research output: Chapter in Book/Report/Conference proceedingChapter

43 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages3-30
Number of pages28
Volume649
DOIs
StatePublished - 2010

Publication series

NameMethods in Molecular Biology
Volume649
ISSN (Print)10643745

Keywords

  • homologous recombination
  • modular assembly
  • single-strand annealing
  • Zinc finger nuclease
  • zinc finger protein engineering methods

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

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    Bhakta, M. S., & Segal, D. (2010). The generation of zinc finger proteins by modular assembly. In Methods in Molecular Biology (Vol. 649, pp. 3-30). (Methods in Molecular Biology; Vol. 649). https://doi.org/10.1007/978-1-60761-753-2_1