A General Strategy for Selecting High-Affinity Zinc Finger Proteins for Diverse DNA Target Sites
- 31 January 1997
- journal article
- other
- Published by American Association for the Advancement of Science (AAAS) in Science
- Vol. 275 (5300), 657-661
- https://doi.org/10.1126/science.275.5300.657
Abstract
A method is described for selecting DNA-binding proteins that recognize desired sequences. The protocol involves gradually extending a new zinc finger protein across the desired 9- or 10-base pair target site, adding and optimizing one finger at a time. This procedure was tested with a TATA box, a p53 binding site, and a nuclear receptor element, and proteins were obtained that bind with nanomolar dissociation constants and discriminate effectively (greater than 20,000-fold) against nonspecific DNA. This strategy may provide important information about protein-DNA recognition as well as powerful tools for biomedical research.Keywords
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