Zinc Finger Consortium: Modular Assembly Kits
Reagents for engineering zinc finger arrays by modular assembly have been described in:
Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly.
Wright et al. Nature Protocols 1, 1637-1652 (2006).
PubMed 17406455, Visit Nature Protocols Website.
UPDATE (May 2008): The laboratories of Keith Joung, Daniel Voytas, and Toni Cathomen have published a large-scale evaluation of the modular assembly method and this report provides important guidance for researchers interested in using this approach to engineer multi-finger domains. The work is described in:
Unexpected failure rates for modular assembly of engineered zinc-fingers.
Ramirez, Foley et al., Nature Methods 5, 374-375.
PubMed 18446154, Visit Nature Methods Website.
IMPORTANT NOTE (July 2012): A legacy version of the ZiFiT software package (ZiFiT v3.3) is 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 at zifit.partners.org.
The legacy version of ZiFiT can be accessed here: ZiFiT v3.3
ZF Consortium Modular Assembly Zinc Finger Set (cat#1000000005)
This kit consists of 141 pc3XB-based zinc finger plasmids encoding 141 different zinc finger modules. ZF1 through ZF57 were derived from the Sangamo module set, ZF58 through ZF106 were derived from the Barbas module set, and ZF107 through ZF141 were derived from the Toolgen module set.
These plasmids are divided into and distributed as two plates: Plate 1, Plate 2.
The information for the 141 zinc finger plasmids is also available as a tab delimited file, which can be opened using Microsoft Excel or a similar spreadsheet application. Download tab delimited file.
The Modular Assembly Zinc Finger Set is $650, plus shipping and handling. This kit will be shipped as bacterial glycerol stocks, 96-well format, on dry ice. Samples should be frozen at -80o C immediately upon arrival. Individual plasmids can be ordered at $65 per plasmid (with discounts for bulk orders), and will be shipped as bacterial stabs.
Note that Addgene uses UPS for shipments and can ship dry ice to most countries, excluding China. Scientists from China interested in obtaining this kit should contact help@addgene.org.
ZF Consortium Modular Assembly Accessory Reagent Set (cat#1000000010)
IMPORTANT NOTE: all of the materials in this accessory kit are included in the ZF Consortium Modular Assembly Zinc Finger Set (above) and therefore do not need to be purchased in addition to the above set.
This set consists of the following five plasmids and one strain that can be used for the bacterial two-hybrid (B2H) system originally described by Hochschild and colleagues (Dove et al., Nature 1997) and later modified by Pabo and colleagues (Joung et al., PNAS 2000). The B2H system is used to screen modularly assembled zinc finger arrays for activity:
The Modular Assembly Accessory Reagent Set is $300 plus shipping and handling, and will be shipped as bacterial stabs at room temperature. Individual plasmids can be ordered for $65 per plasmid, and will be shipped as bacterial stabs as well.