title: TALEN and CRISPR/Cas9-mediated genome editing in the early-branching metazoan Nematostella vectensis.
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.
authors.affiliation: Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, Missouri 64110, USA.
authors.forename: Aissam
authors.initials: A
authors.lastname: Ikmi
authors.affiliation: Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, Missouri 64110, USA.
authors.forename: Sean A
authors.initials: SA
authors.lastname: McKinney
authors.affiliation: Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, Missouri 64110, USA.
authors.forename: Kym M
authors.initials: KM
authors.lastname: Delventhal
authors.affiliation: 1] Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, Missouri 64110, USA [2] Department of Anatomy and Cell Biology, University of Kansas School of Medicine, Kansas City, Kansas 66160, USA.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Gibson
mesh_terms: D000818:Animals; D001483:Base Sequence; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D003851:Deoxyribonucleases / Q000378:metabolism*; D004720:Endonucleases / Q000378:metabolism*; D016678:Genome; D059765:Homologous Recombination; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012615:Sea Anemones / Q000235:genetics* / Q000254:growth & development
chemical_list: D003851:Deoxyribonucleases; D004720:Endonucleases