Wolfe-Lawson ZFN modular assembly kit
Zinc-finger nucleases (ZFNs) allow targeted gene inactivation in a wide range of model organisms. We provided an archive of 81 clones containing 27 different zinc-finger modules at finger 1, 2 and 3 positions for straightforward construction of ZFNs using modular assembly-based approach. The majority of zinc-finger proteins assembled from these modules have favorable specificities and nearly one-third of modular ZFNs generated lesions at their targets in the zebra fish genome. These modules are designed for the construction of three-finger zinc-finger ZFNs, but they can also be used for assembling longer ZFNs containing 4 or more modules. We set up a website to help users search appropriate sites in their target sequences and design corresponding ZFNs using this archive:
To facilitate the application of ZFNs within the zebrafish community we constructed a public database of sites in the zebrafish genome that can be targeted using this archive:
Protocols for modular assembly construction of ZFNs can be found at the above website as well as in the following publication:
Evaluation and application of modularly assembled zinc finger nucleases in zebrafish. Zhu C, Smith T, McNulty J, Rayla AL, Lakshmanan A, Siekmann AF, Buffardi M, Meng X, Shin J, Padmanabhan A, Cifuentes D, Giraldez AJ, Look AT, Epstein JA, Lawson ND, and Wolfe SA. Development, 2011, 138:4555-4564. PubMed 21937602.
Figure: Modules containing in this archive can recognize the following triplet target sites (5’→3’):
The kit is $350, plus shipping and handling. This kit consists of 1 plate, and will be shipped as bacterial glycerol stocks, 96-well format, on dry ice. Samples should be frozen at -80 degrees Celsius immediately upon arrival. Individual plasmids can be ordered at $65 per plasmid, and will be shipped as bacterial stabs.
Download Information and Plate Map PDF (included with shipment).
The kit consists of 71 plasmids provided in a single 96-well plate.
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 firstname.lastname@example.org.