Patent ID: 7241573
Filing Date: 2007-07-10
Classification: C07K,C12N,C40B

Abstract:
1. A method for binding a nucleic acid binding protein to a target nucleotide sequence, wherein the binding protein comprises a plurality of zinc fingers, and further wherein at least two 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 G, then position +6 in the α-helix is Arg or Lys; b) if base 4 in the quadruplet is A, then position +6 in the α-helix is Glu, Asn or Val; c) if base 4 in the quadruplet is T, then position +6 in the α-helix is Ser, Thr, Val or Lys; d) if base 4 in the quadruplet is C, then position +6 in the α-helix is Ser, Thr, Val, Ala, Glu or Asn; e) if base 3 in the quadruplet is G, then position +3 in the α-helix is His; f) if base 3 in the quadruplet is A, then position +3 in the α-helix is Asn; g) if base 3 in the quadruplet is T, then position +3 in the α-helix is Ala, Ser or Val; h) if base 3 in the quadruplet is C, then position +3 in the α-helix is Ser, Asp, Glu, Leu, Thr or Val; i) if base 2 in the quadruplet is G, then position −1 in the α-helix is Arg; j) if base 2 in the quadruplet is A, then position −1 in the α-helix is Gln; k) if base 2 in the quadruplet is T, then position −1 in the α-helix is His or Thr; l) if base 2 in the quadruplet is C, then position −1 in the α-helix is Asp or His; m) if base 1 in the quadruplet is G, then position +2 is Glu; n) if base 1 in the quadruplet is A, then position +2 Arg or Gln; o) if base 1 in the quadruplet is C, then position +2 is Asn, Gln, Arg, His or Lys; p) if base 1 in the quadruplet is T, then position +2 is Ser or Thr; and the method comprises (iii) preparing a nucleic acid encoding the binding protein of (ii); and (iv) introducing the nucleic acid of (iii) into a cell, whereby the encoded nucleic acid binding protein is expressed and binds to the target sequence.