Golden Gate TALEN and TAL Effector Kit 2.0
TALENs are fusions of transcription activator-like (TAL) effectors of plant pathogenic Xanthomonas spp. to the FokI nuclease. TALENs bind and cleave DNA in pairs, and their binding specificity is determined by customizable arrays of polymorphic amino acid repeats in the TAL effectors. This collection and accompanying documentation allow one to efficiently assemble TALEN constructs with custom repeat arrays, containing anywhere between 12 and 31 repeats.
This kit includes 14 new plasmids plus the initial plasmids described in:
Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Apr 14. PUBMED. Open Access Article.
Click here for a description of the new plasmids included in Golden Gate TALEN and TAL Effector Kit 2.0
In January 2013, additional plasmids were added to the Golden Gate kit. They include the 11 NH RVD module plasmids. Streubel et al. (2012) and Cong et al. (2012) show that the RVD NH targets G more specifically than NN does, with good apparent affinity. NK also targets G specifically, but with lower affinity (Christian et al., 2012). NH is therefore the currently preferred RVD for targeting G.
pZHY500 and pZHY501 are Golden Gate compatible yeast expression vectors with deletions in the coding sequence that result in truncations of both the amino and carboxyl portions of the TAL portion of the protein to match the "Miller et al., 2011" architecture. Details can be found in Plant Physiol. 2013 Jan;161(1):20-7.
pCP5b is an unpublished plasmid that allows one to test TALEN activity in yeast. This plasmid is very similar to and can be used in place of the published pCP5 (see Proc Natl Acad Sci U S A. 107(26):12028-33). You can find a detailed protocol on how to design and clone targets for the Yeast TALEN assay HERE.
Web based software to search for TAL binding sites
Download a ZIP file containing all plasmid sequences (except pKEB31) in GenBank or .txt format and all plasmid maps in JPEG or PDF format
The kit is $425, plus shipping and handling. This kit will be shipped to most countries as bacterial glycerol stocks, 96-well format, on dry ice. Samples should be frozen at -80°C immediately upon arrival. Scientists in countries that cannot receive dry ice shipments will receive a room temperature version of the kit that needs to be processed immediately upon arrival. Individual plasmids can be ordered at $65 per plasmid, and will be shipped as bacterial stabs.