TALEN construction via “Unit Assembly” method and targeted genome modifications in zebrafish

Transcription activator-like effector nucleases (TALENs) are engineered endonucleases composed of a customized transcription activator-like effector (TALE) DNA-binding domain and a FokI DNA cleavage domain. TALENs induce DNA double-strand breaks (DSBs) at their target sites on the chromosome and have been successfully used for genome engineering in many species and cultured cells. Zebrafish is a very popular model organism in both basic and clinical research. Here, we describe the details of construction of customized TALENs using the “Unit Assembly” (UA) method, as well as three applications of zebrafish genome manipulations using TALENs: gene knock-out, large chromosome deletion, and gene knock-in by homologous recombination.