TALEN and CRISPR/Cas9-mediated genome editing in the early-branching metazoan Nematostella vectensis

Nat Commun. 2014 Nov 24;5:5486. doi: 10.1038/ncomms6486.

Abstract

Non-bilaterian phyla represent key lineages for exploring the evolutionary history of early animals. However, despite an increasing number of sequenced genomes from early-branching metazoans, efficient and reproducible methodologies for analysis of gene function remain a major challenge. Here we report the utilization of the TALEN and CRISPR/Cas9 systems to induce targeted mutations and homologous recombination-mediated transgenesis in the sea anemone Nematostella vectensis. We also present a new method to isolate genetically modified animals using engineered selection cassettes introduced by homologous recombination. Taken together, these methods will permit sophisticated gain- and loss-of-function analyses in Nematostella and perhaps other early metazoan species that allow for zygotic injection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Deoxyribonucleases / metabolism*
  • Endonucleases / metabolism*
  • Genome*
  • Homologous Recombination
  • Molecular Sequence Data
  • Mutagenesis
  • Sea Anemones / genetics*
  • Sea Anemones / growth & development

Substances

  • Deoxyribonucleases
  • Endonucleases