Peptides and uses
View Patent ↗Disclosed herein are zinc finger peptides having at least 4 zinc finger domains, such as 6, 11, 12 or 18 zinc finger domains. The zinc finger peptides may be fused to effector domains for modulating gene expression. Zinc finger peptides of the invention are useful for targeting trinucleotide-repeat sequences in mutant genes and may have applications in gene therapy. The zinc finger peptides may have nucleic acid recognition sequences according to SEQ ID NO: 101. Also disclosed are methods for constructing poly-zinc finger peptides having arrays of at least 8 zinc finger domains, along with zinc finger frameworks that may be useful for selecting zinc finger peptides from libraries.
1. A zinc finger peptide comprising the sequence:
N′-[(Formula 2)-X 6 ] n0 -{[(Formula 2)-X 5 -(Formula 2)-X 6 ] n1 -[(Formula 2)-X5-(Formula 2)-X L ]} n2 -[(Formula 2)-X 5 -(Formula 2)-X 6 ] n3 -[(Formula 2)-X 5 -(Formula 2)]-[X 6 -(Formula 2)-] n4 -C′,
wherein n0 is 0 or 1, n1 is from 1 to 4, n2 is 1 or 2, n3 is from 1 to 4, n4 is 0 or 1, X 5 is a linker sequence of 5 amino acids, X 6 is a linker sequence of 6 or 7 amino acids, and X L is a linker sequence of at least 8 amino acids, and Formula 2 is a zinc finger domain of the sequence -X 0-2 CX 1-5 CX 2-7 X −1 x +1 x +2 x +3 x +4 x +5 x +6 HX 3-6 H / C -, wherein X is any amino acid, the numbers in subscript indicate the possible numbers of residues represented by X at that position, and the number in superscript indicates the position of the amino acid in the recognition sequence of the zinc finger domain;
wherein the zinc finger peptide comprises from 8 to 32 zinc finger domains,
comprises at least the sequence of SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 108 or a sequence having at least 90% sequence identity thereto, and
at least one zinc finger domain comprises a X −1 X +1 X +2 X +3 X +4 X +5 X +6 recognition sequence according to SEQ ID NO: 101.
2. The zinc finger peptide of claim 1 , which comprises 10, 11, 12 or 18 zinc finger domains.
3. The zinc finger peptide of claim 1 , wherein at least one X −1 X +1 X +2 X +3 X +4 X +5 X +6 sequence is the amino acid sequence QRATLQR (SEQ ID NO: 1).
4. The zinc finger peptide of claim 1 , which binds double-stranded trinucleotide repeat nucleic acid sequences comprising CAG-repeat, CTG-repeat, and/or CAGCTG-repeat sequences containing at least 22, at least 35 or at least 63 contiguous triplet repeats.
5. The zinc finger peptide of claim 1 , which binds double-stranded trinucleotide repeat nucleic acid sequences containing at least 22, at least 35 or at least 63 contiguous triplet repeats, and wherein the binding affinity is at least 1 nM; at least 100 pM or at least 10 pM.
6. The zinc finger peptide of claim 1 , which is fused to and at least one effector domain, selected from transcriptional repressor domains, transcriptional activator domains, transcriptional insulator domains, chromatin remodelling, condensation or decondensation domains, nucleic acid or protein cleavage domains, dimerisation or multimermisation domains, enzymatic domains, signalling/targeting sequences or domains.