Genome engineering in mice using TALEN or CRISPR/Cas9 nucleases.

In the course of this work, TALENs were optimized to increase the rate and specificity of site-directed gene modifications. Furthermore, mice encoding genomic deletions of 9.3 kb or 3.2 kb in Rab38 could be generated at a frequency of 6-37% through pronuclear microinjection of TALEN or CRISPR/Cas9 nuclease pairs. Application of the CRISPR/Cas9 system led to deletions of up to 52 kb in vitro. This work shows that even the homozygous excision of large gene segments can be obtained in a single step and contributes to the investigation of human hereditary diseases