Design of polydactyl zinc-finger proteins for unique addressing within complex genomes

Qiang Liu, David Segal, Jayant B. Ghiara, Carlos F. Barbas

Research output: Contribution to journalArticle

211 Scopus citations

Abstract

Zinc-finger proteins of the Cys2-His2 type represent a class of malleable DNA-binding proteins that may be selected to bind diverse sequences. Typically, zinc-finger proteins containing three zinc-finger domains, like the murine transcription factor Zif268 and the human transcription factor Sp1, bind nine contiguous base pairs. To create a class of proteins that would be generally applicable to target unique sites within complex genomes, we have utilized structure-based modeling to design a polypeptide linker that fuses two three-finger proteins. Two six-fingered proteins were created and demonstrated to bind 18 contiguous bp of DNA in a sequence-specific fashion. Expression of these proteins as fusions to activation or repression domains allows transcription to be specifically up- or down-modulated within human cells. Polydactyl zinc-finger proteins should be broadly applicable as genome-specific transcriptional switches in gene therapy strategies and the development of novel transgenic plants and animals.

Original languageEnglish (US)
Pages (from-to)5525-5530
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number11
DOIs
StatePublished - May 27 1997
Externally publishedYes

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Keywords

  • Gene therapy
  • Genome addressing
  • Molecule design
  • Transcriptional regulation
  • Zinc-finger proteins

ASJC Scopus subject areas

  • Genetics
  • General

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