Improved DNA binding specificity from polyzinc finger peptides by using strings of two-finger units

Multizinc finger peptides are likely to reach increased prominence in the s earch for the "ideal" designer transcription factor for in vivo application s such as gene therapy. However, for these treatments to be effective and s afe, the peptides must bind with high affinity and, more importantly, with great specificity. Our previous research has shown that zinc finger arrays can be made to bind 18 bp of DNA with picomolar affinity, but also has sugg ested that arrays of fingers also may bind tightly to related sequences. Th is work addresses the question of zinc finger DNA binding specificity. We s how that by changing the way in which zinc finger arrays are constructed-by linking three two-finger domains rather than two three-finger units-far gr eater target specificity can be achieved through increased discrimination a gainst mutated or closely related sequences. These new peptides have the ad ded capability of being able to span two short gaps of unbound DNA, althoug h still binding with picomolar affinity to their target sites. We believe t hat this new method of constructing zinc finger arrays will offer greater e fficacy in the fields of gene therapy and in the production of transgenic o rganisms than previously reported zinc finger arrays.