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Golden gate talen kit v2

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The Golden Gate TALEN kit v2.0 is a tool designed for the assembly of Transcription Activator-Like Effector Nucleases (TALENs). TALENs are a type of genome editing technology used to modify targeted DNA sequences. The kit provides the necessary components and protocols for the construction of custom TALEN proteins.

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Close spelling variants of this product

Golden Gate TALEN kit (10 citations) Golden Gate TALEN (5 citations) Golden Gate kit (3 citations) TALENs (2 citations)

2 protocols using golden gate talen kit v2

1

TALEN Mutagenesis of eng Gene

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eng mutants were generated by TALEN mutagenesis. TALENs specific to eng were designed using the online TAL Effector Nucleotide Targeter (https://talent.cac.cornell.edu/) with the 1st exon provided as target sequence. Criteria for optimal TALEN pairs was followed as described in Cermak et al. 2011. The following TALEN pair was selected for optimal length and presence of an MspI site in the spacer, facilitating identification of mutations by PCR and digest.
5’ TALEN armDNA sequence: 5’-tgctgtgtttgcttct-3’
RVD sequence: NN HD NG NN NG NN NG NG NG NN NI HD NG NG HD NG
3’ TALEN armDNA sequence: 5’-tggagctcttacctga-3’
RVD sequence: NN NN NI NN HD NG HD NG NG NI HD HD NG NN NI
RVDs were assembled using the Golden Gate TALEN kit v2.0 (Addgene), into pTAL4 vectors
Sugden W.W., Meissner R., Aegerter-Wilmsen T., Tsaryk R., Leonard E.V., Bussmann J., Hamm M.J., Herzog W., Jin Y., Jakobsson L., Denz C, & Siekmann A.F. (2017). Endoglin controls blood vessel diameter through endothelial cell shape changes in response to haemodynamic cues. Nature cell biology, 19(6), 653-665.
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2

Generating Engineered eng Mutants

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eng mutants were generated by TALEN mutagenesis. TALENs specific to eng were designed using the online TAL Effector Nucleotide Targeter (https://talent.cac.cornell.edu/) with the 1st exon provided as target sequence. Criteria for optimal TALEN pairs was followed as described in Cermak et al. 2011. The following TALEN pair was selected for optimal length and presence of an MspI site in the spacer, facilitating identification of mutations by PCR and digest.
5’ TALEN armDNA sequence: 5’-tgctgtgtttgcttct-3’
RVD sequence: NN HD NG NN NG NN NG NG NG NN NI HD NG NG HD NG
3’ TALEN armDNA sequence: 5’-tggagctcttacctga-3’
RVD sequence: NN NN NI NN HD NG HD NG NG NI HD HD NG NN NI
RVDs were assembled using the Golden Gate TALEN kit v2.0 (Addgene), into pTAL4 vectors
Sugden W.W., Meissner R., Aegerter-Wilmsen T., Tsaryk R., Leonard E.V., Bussmann J., Hamm M.J., Herzog W., Jin Y., Jakobsson L., Denz C, & Siekmann A.F. (2017). Endoglin controls blood vessel diameter through endothelial cell shape changes in response to haemodynamic cues. Nature cell biology, 19(6), 653-665.
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