An improved strategy for constructing "designer" Cys2His2 zinc finger proteins.
Extract: The Cys2His2 zinc finger domain (hereafter simply "zinc finger") provides a useful scaffold for creating customized DNA-binding proteins, a technology with potential applications in biological research, molecular medicine, and gene therapy. Using a combination of targeted randomization and selection methodologies (e.g., phage display), many research groups have successfully altered the DNA-binding specificities of single zinc fingers, which typically recognize three to four base pairs of DNA. In these experiments, potential DNA-binding residues in a finger's a-helix (or "recognition helix") were randomized to generate a library of variants and then selection methods were used to identify fingers with desired DNA-binding specificities. To create synthetic multi-finger proteins capable of recognizing longer DNA sequences, various investigators have linked together three or more pre-selected or pre-characterized finger domains. This type of strategy, which assumes that individual fingers behave in a modular fashion, permits the rapid assembly of multi-finger proteins directed to bind a wide variety of different DNA sequences.
Thibodeau
Stacey A
SA
Molecular Pathology Unit, Division of Molecular Pathology and Research, Department of Pathology, Massachusetts General Hospital, 149 13th Street, Charlestown, MA 02129, USA.
Joung
J Keith
JK