Patent ID: 6746838
Filing Date: 2004-06-08
Classification: C07K,C12N,C40B

Abstract:
A method for preparing a nucleic acid binding protein that binds to a target nucleotide sequence, wherein the binding protein comprises a plurality of zinc fingers of the Cys2-His2 class, further wherein adjacent zinc fingers bind synergistically to overlapping quadruplet target subsites, wherein the method comprises:i) selecting a quadruplet within the target nucleotide sequence; ii) designing the binding protein such that binding of a zinc finger to the quadruplet is obtained by choosing the sequence of particular residues of the zinc finger depending on the nucleotide sequence of the quadruplet, as follows: a) if base 4 in the quadruplet is A, then position +6 in the &agr;-helix is Glu, Asn or Val; b) if base 4 in the quadruplet is C, then position +6 in the &agr;-helix is Ser, Thr, Val, Ala, Glu or Asn c) if base 4 in the quadruplet is G, then position +6 in the &agr;-helix is Arg or Lys; d) if base 4 in the quadruplet is T, then position +6 in the &agr;-helix is Ser, Thr, Val or Lys; iii) synthesizing a polynucleotide encoding the binding protein of (ii); iv) introducing the polynucleotide of (iii) into a cell; and v) incubating the cell under conditions in which the encoded nucleic acid binding protein is expressed.