A GENERAL STRATEGY FOR SELECTING HIGH-AFFINITY ZINC-FINGER PROTEINS FOR DIVERSE DNA TARGET SITES

A method is described for selecting DNA-binding proteins that recogniz e desired sequences. The protocol involves gradually extending a new z inc 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, a nd proteins were obtained that bind with nanomolar dissociation consta nts and discriminate effectively (greater than 20,000-fold) against no nspecific DNA. This strategy may provide important information about p rotein-DNA recognition as well as powerful tools for biomedical resear ch.