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

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS

卷:19

Article

Segal, David J.¹ 

[1] Univ Calif Davis, Genome Ctr, Davis, CA 95616 USA

关键词: Gene therapy;    Gene targeting;    Homologous recombination;    Targeted mutagenesis;    Protein engineering;    Zinc finger nuclease;   

DOI  :  10.1016/j.bmcl.2009.02.109

来源: Elsevier

【 摘 要 】

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. (C) 2009 Elsevier Ltd. All rights reserved.

【 授权许可】

Free