A General Strategy for Selecting High-Affinity Zinc Finger Proteins for Diverse DNA Target Sites
@article{Greisman1997AGS, title={A General Strategy for Selecting High-Affinity Zinc Finger Proteins for Diverse DNA Target Sites}, author={Harvey A. Greisman and Carl O. Pabo}, journal={Science}, year={1997}, volume={275}, pages={657 - 661} }
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…
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