TALEN-Mediated Mutagenesis and Genome Editing

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1451; p. 17
• Main Authors: Ma, Alvin C H, Chen, Yi, Blackburn, Patrick R, Ekker, Stephen C
• Format: Journal Article
• Language: English
• Published: United States 2016
• Subjects: Animals; CRISPR-Cas Systems - genetics; DNA Breaks, Double-Stranded; Gene Editing - methods; Mutagenesis - genetics; Transcription Activator-Like Effector Nucleases - genetics; Zebrafish - genetics; Zebrafish; Customized nucleases; Genome editing; Golden Gate; FusX; TALEN
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-3771-4_2

Transcription activator-like effectors (TALEs) are important genomic tools with customizable DNA-binding motifs for locus-specific modifications. In particular, TALE nucleases or TALENs have been successfully used in the zebrafish model system to introduce targeted mutations via repair of double-stranded breaks (DSBs) either through nonhomologous end joining (NHEJ) or by homology-directed repair (HDR) and homology-independent repair in the presence of a donor template. Compared with other customizable nucleases, TALENs offer high binding specificity and fewer sequence constraints in targeting the genome, with comparable mutagenic activity. Here, we describe a detailed in silico design tool for zebrafish genome editing for TALENs and CRISPR/Cas9 custom restriction enzymes using Mojo Hand 2.0 software.