Genome editing using artificial site-specific nucleases in zebrafish

Dev Growth Differ. 2014 Jan;56(1):26-33. doi: 10.1111/dgd.12094. Epub 2013 Oct 14.

Abstract

Zebrafish is a model vertebrate suitable for genetic analysis. Forward genetic analysis via chemical mutagenesis screening has established a variety of zebrafish mutants that are defective in various types of organogenesis, and the genes responsible for the individual mutants have been identified from genome mapping. On the other hand, reverse genetic analysis via targeted gene disruption using embryonic stem (ES) cells (e.g., knockout mouse) can uncover gene functions by investigating the phenotypic effects. However, this approach is mostly limited to mice among the vertebrate models because of the difficulty in establishing ES cells. Recently, new gene targeting technologies, such as the transcription activator-like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 systems, have been developed: that can directly introduce genome modifications at the targeted genomic locus. Here, we summarize these new and powerful genome editing techniques for the study of zebrafish.

Keywords: clustered regularly interspaced short palindromic repeats/Cas9; genome editing; heteroduplex mobility assay; transcription activator-like effector nucleases.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • CRISPR-Cas Systems
  • DNA Restriction Enzymes / genetics
  • DNA Restriction Enzymes / metabolism*
  • Genetic Engineering / methods*
  • Genome / genetics*
  • Substrate Specificity
  • Zebrafish / genetics*

Substances

  • DNA Restriction Enzymes