TALEN-Mediated Mutagenesis and Genome Editing

Methods Mol Biol. 2016:1451:17-30. doi: 10.1007/978-1-4939-3771-4_2.

Abstract

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.

Keywords: Customized nucleases; FusX; Genome editing; Golden Gate; TALEN; Zebrafish.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • DNA Breaks, Double-Stranded
  • Gene Editing / methods*
  • Mutagenesis / genetics*
  • Transcription Activator-Like Effector Nucleases / genetics*
  • Zebrafish / genetics

Substances

  • Transcription Activator-Like Effector Nucleases