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Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys2-His2 Zinc-finger Domains
Published on: March 25, 2015
Engineering Cys2His2 zinc finger domains using a bacterial cell-based two-hybrid selection system
Stacey Thibodeau-Beganny1, J Keith Joung
1Massachusetts General Hospital, Charlestown, USA.
Methods in Molecular Biology (Clifton, N.J.)
|March 5, 2008
Summary
This study details updated protocols for bacterial two-hybrid systems, a highly effective method for selecting novel synthetic Cys2His2 zinc finger domains with specific DNA-binding capabilities from large libraries.
Area of Science:
- Molecular Biology
- Protein Engineering
- Biotechnology
Background:
- Synthetic Cys2His2 zinc finger domains offer customizable DNA-binding specificities.
- Selection methodologies like phage display are used to identify these domains from randomized libraries.
- Bacterial cell-based two-hybrid systems have proven effective for zinc finger selection.
Purpose of the Study:
- To provide updated and detailed protocols for performing zinc finger selections.
- To highlight the utility of the bacterial two-hybrid system for this purpose.
Main Methods:
- Utilizing a bacterial cell-based two-hybrid system for selection.
- Employing large randomized libraries of Cys2His2 zinc finger domains.
Main Results:
- The bacterial two-hybrid system is demonstrated to be effective for selecting zinc fingers with desired specificities.
- Detailed protocols are presented for implementing this selection methodology.
Conclusions:
- The bacterial two-hybrid system is a robust and efficient method for identifying novel synthetic zinc finger domains.
- Updated protocols facilitate the application of this system in protein engineering and synthetic biology research.
