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Engineered Zinc Finger Proteins

Volume 649 of the series Methods in Molecular Biology pp 31-50

Date:

Engineering Single Cys2His2 Zinc Finger Domains Using a Bacterial Cell-Based Two-Hybrid Selection System

  • Stacey Thibodeau-BegannyAffiliated withMolecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital
  • , Morgan L. MaederAffiliated withMolecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General HospitalBiological and Biomedical Sciences Program, Harvard Medical School
  • , J. Keith JoungAffiliated withMolecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General HospitalBiological and Biomedical Sciences Program and Department of Pathology, Harvard Medical School

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Abstract

Individual synthetic Cys2His2 zinc finger domains with novel DNA-binding specificities can be identified from large randomized libraries using selection methodologies such as phage display. We have previously demonstrated that a bacterial cell-based two-hybrid system is at least as effective as phage display for selecting zinc fingers with desired specificities from such libraries. In this chapter we provide updated, detailed protocols for performing zinc finger selections using the bacterial two-hybrid system.

Key words

Bacterial two-hybrid zinc finger selection