Zinc Finger Consortium - Modular Assembly Kits
(Kit # 1000000005, 1000000010 )

Original Publication

Description

Zinc fingers are the most common family of transcription factors in organisms ranging from yeast to humans. Custom zinc finger proteins can be designed to target a specific DNA sequence in a gene of interest in nearly any cell type.

Zinc finger nucleases (ZFNs), which consist of an engineered zinc finger DNA-binding domain fused to the DNA cleavage domain of the FokI enzyme, are powerful tools for performing genome engineering in a variety of organisms. ZFNs can introduce double-stranded breaks in DNA that stimulate both homologous and non-homologous recombination pathways to alter a desired locus in the genome.

The zinc finger modules in this kit were derived from the Barbas, Sangamo and ToolGen module sets and standardized to permit "mixing-and-matching" of the different modules. A complementary web-based Zinc Finger Targeter program (ZiFiT) was designed to assist in the identification of potential DNA target sites.

In May 2008, the laboratories of Keith Joung, Daniel Voytas, and Toni Cathomen published a large-scale evaluation of the modular assembly method and reported important guidance for researchers interested in using this approach to engineer multi-finger domains. The work is described in:

Kit Documentation

Links

In July 2012, a legacy version of the ZiFiT software package (ZiFiT v3.3) was made available to maintain support for modular assembly of zinc fingers using the Modular Assembly Kit developed by the Zinc Finger Consortium (Wright et al., Nature Protocols 1, 1637-1652, 2006). Considerable progress has been made towards engineering custom DNA binding proteins and the Zinc Finger Consortium now strongly recommends utilizing newer alternative methods for engineering DNA binding domains. For more information on these methods please see the current version of ZiFiT.

Resources

A spreadsheet containing information for the 141 zinc finger plasmids is available as a CSV file:

How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:
"The plasmid kit used for engineering ZFNs was a gift from Keith Joung and Dan Voytas (Addgene kit #1000000005 or #1000000010 )"

For your Reference section:
Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly. Wright DA, Thibodeau-Beganny S, Sander JD, Winfrey RJ, Hirsh AS, Eichtinger M, Fu F, Porteus MH, Dobbs D, Voytas DF, Joung JK. Nat Protoc . Nat Protoc. 2006;1(3):1637-52. PubMed 17406455.