Restricted spacer tolerance of a zinc finger nuclease with a six amino acid linker

Yuka Shimizu, Mital S. Bhakta, David Segal

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Zinc finger nucleases can be engineered to create highly efficient and precise changes to the genetic information within living cells. We report the investigation of an important parameter that defines the type of target site the nuclease can cleave. The active nuclease is a dimer, requiring that the DNA target site contain two zinc finger binding sites separated by a short spacer. Using a plasmid-based recombination assay in HEK 293T cells, we show that a 6 amino acid linker between the zinc finger DNA-binding domain and the FokI cleavage domain restricts nuclease activity to sites containing a 6 bp spacer. These observations concur with other recent studies, suggesting this information will be useful in the design of new potent and accurate zinc finger nucleases.

Original languageEnglish (US)
Pages (from-to)3970-3972
Number of pages3
JournalBioorganic and Medicinal Chemistry Letters
Volume19
Issue number14
DOIs
StatePublished - Jul 15 2009

Fingerprint

Zinc Fingers
Zinc
Amino Acids
HEK293 Cells
DNA
Dimers
Genetic Recombination
Assays
Plasmids
Binding Sites
Cells

Keywords

  • Gene targeting
  • Gene therapy
  • Homologous recombination
  • Protein engineering
  • Targeted mutagenesis
  • Zinc finger nuclease

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Drug Discovery
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology
  • Clinical Biochemistry
  • Biochemistry

Cite this

Restricted spacer tolerance of a zinc finger nuclease with a six amino acid linker. / Shimizu, Yuka; Bhakta, Mital S.; Segal, David.

In: Bioorganic and Medicinal Chemistry Letters, Vol. 19, No. 14, 15.07.2009, p. 3970-3972.

Research output: Contribution to journalArticle

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