Open AccessImproved DNA binding specificity from polyzinc finger peptides by using strings of two-finger units
Author(s) -
Michael J. Moore,
A. Klug,
Yen Choo
Publication year - 2001
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.98.4.1437
Subject(s) - zinc finger , zinc finger nuclease , dna , ring finger domain , computational biology , dna binding protein , sp1 transcription factor , gene , biology , lim domain , transcription factor , binding selectivity , dna sequencing , biochemistry , promoter , gene expression
Multizinc finger peptides are likely to reach increased prominence in the search for the “ideal” designer transcription factor forin vivo applications such as gene therapy. However, for these treatments to be effective and safe, 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 suggested that arrays of fingers also may bind tightly to related sequences. This work addresses the question of zinc finger DNA binding specificity. We show 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 greater target specificity can be achieved through increased discrimination against mutated or closely related sequences. These new peptides have the added capability of being able to span two short gaps of unbound DNA, although still binding with picomolar affinity to their target sites. We believe that this new method of constructing zinc finger arrays will offer greater efficacy in the fields of gene therapy and in the production of transgenic organisms than previously reported zinc finger arrays.