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.
IMPORTANT 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.
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.
Browse zinc finger plasmids by target DNA sequence and finger amino acid sequence.
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.