TALENs

TALEN-based engineering at PolyGene.

PolyGene offers services based on Transcription Activator-Like Effector Nuclease (TALEN) technology for the speedy generation of constitutive knockout mice, as well as rats and rabbits.

TALEs (Transcription Activator-Like Effectors) are proteins that bind to DNA in a sequence-specific way (Sun and Zhao, 2013). The TALEs are constructed from modularly organized DNA-binding domains, tested and validated in vitro. By fusing such a TALE to a Nuclease (a “TALEN”), a highly specific DNA scissor is made.

talens_mouse_explained
A TALEN acts as a heterodimer (two units of a TALE DNA binding domain (brown, orange) fused to a catalytic domain (grey)), cleaving two close sequences, and thereby increasing specificity to a complexity of 30 – 40 base pairs. Within the cells, TALEN generate double strand breaks which are repaired by Non-Homologous End Joining (NHEJ). NHEJ is, a natural but error prone repair mechanism that can be used to introduce nucleotide deletions or insertions. This can lead to inactivation or knockout of a target gene.

Within the cells, TALEN generate double strand breaks which are repaired by Non-Homologous End Joining (NHEJ). NHEJ is, a natural but error prone repair mechanism that can be used to introduce nucleotide deletions or insertions. This can lead to inactivation or knockout of a target gene.

Sun N, Zhao H.
Transcription activator-like effector nucleases (TALENs): a highly efficient and versatile tool for genome editing.
Biotechnol Bioeng. 2013; 110(7):1811-21.

Advantages
The use of a pair of TALE molecules with two recognition sites of 16-18 bp each makes this system very specific, generating only very few off-target effects.
Disadvantages
The construction of the large TALEN molecules is more time consuming and more restricted in the choice of targets than for the CRISPR/Cas9 system. The efficiency of TALEN-mediated knockout is lower than for CRISPR/Cas9 allowing mainly for the generation of knockout animals. For knock-in generation, the CRISPR/Cas9 system is better suited.
For the generation of other TALE fusion proteins, such as sequence specific activators or silencers, please inquire.

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