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A General Strategy for Selecting High-Affinity Zinc Finger Proteins for Diverse DNA Target Sites

Science  31 Jan 1997:
Vol. 275, Issue 5300, pp. 657-661
DOI: 10.1126/science.275.5300.657

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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.

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