Conference on Transposition and Genome Engineering

Come visit us at the booth!

Don't miss our oral presentation on September 20th:

The next generation of TALEN™ tools for efficient cell engineering
Dr Alexandre Juillerat

The ability to disrupt or modify cellular functions through accurate and easy addition, removal, or exchange of DNA sequences directly in cellular genomes, together with the advances in, and access to, genome sequencing and epigenetic information, is opening exciting new perspectives in biology, biotechnology and medicine. Over the last few years, the transcription activator-like effector (TALE) scaffold has proven to be a versatile and incredibly robust platform for generating transcription activators, repressors, recombinases, and nucleases with custom DNA specificity. Here we present our platform allowing high throughput production of TALEN™. Based on our nuclease activity assays, we demonstrate how to ensure the specificity of a TALEN™ and its accessibility to methylated loci by using alternative modules, such as N*. We illustrate the use of TALEN™ tools for different applications, employing the non-homologous end-joining pathway for knock-out, and homologous recombination for gene tagging and nucleotide substitution. Until now, TALEN™ have been designed according to a unique dimeric architecture that uses FokI as nuclease domain. Generating new architectures that do not require dimerization would enlarge the scope of applications for TALEN™. The modularity of the TALE scaffold opens the possibility to further engineer TALE-based molecular tools. Based on this, we have developed a new TALE scaffold to create a novel class of monomeric, DNA-cleaving enzymes known as Compact TALEN™. In vivo experiments in various organisms have demonstrated that the half-size, single-polypeptide Compact TALEN™ exhibit overall activity in line with the current classical TALEN™ designer nucleases.