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Voytas Lab - Golden Gate TALEN and TAL Effector Kit 1.0
(Kit #  Discontinued )

Depositing Lab:   Daniel Voytas

This kit has been discontinued. The newest version (v 2.0) is available here.

The TALEN plasmids fill 72 wells on a single 96-well plate.

Original Publication

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 21493687

Description

TALENs are important new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors of plant pathogenic Xanthomonas spp. to the FokI nuclease, TALENs bind and cleave DNA in pairs. 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. The reagents include a plasmid construct for making custom TAL effectors and one for TAL effector fusions to additional proteins of interest. The web based software which allows one to search for TAL binding sites is located at: http://boglabx.plp.iastate.edu/TALENT/

Click here to see available add-on plasmids generated for use with this kit

Kit Documentation

Icon Golden Gate plate map (1.2 MB)

Resources

Icon Voytas GG TALEN Kit Maps Sequences (4.3 MB)

Protocols

Icon Golden Gate assembly protocol (637.1 KB)

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 generation of TALENs was a gift from Daniel Voytas (Addgene kit # 1000000016)"

For your Reference section:

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 21493687

Golden Gate TALEN and TAL Effector Kit 1.0 supplemental plasmids

These 14 plasmids are available in the Golden Gate TALEN and TAL Effector Kit 2.0, but were not included in the original (v 1.0) 72 plasmid version of the kit.

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). Read the detailed protocol on how to design and clone targets for the Yeast TALEN assay.

Updated protocols, maps, and references can be found on the Golden Gate TALEN and TAL Effector Kit 2.0 page.

IMPORTANT NOTE: Like the pNK series of plasmids, several of the pNH plasmids have varying lengths that differ from what is reported on the maps. The size discrepancies do not appear to affect function. Addgene does not recommend verifying these plasmids via diagnostic digest.

Plasmid Resistance
pZHY500 Ampicillin
pZHY501 Ampicillin
pCP5b Ampicillin
pLR-NH Spectinomycin
pNH1 Tetracycline
pNH2 Tetracycline
pNH3 Tetracycline
pNH4 Tetracycline
pNH5 Tetracycline
pNH6 Tetracycline
pNH7 Tetracycline
pNH8 Tetracycline
pNH10 Tetracycline
pNH9 Tetracycline
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