title: Tissue-specific genome editing in Ciona embryos by CRISPR/Cas9.
abstract: The CRISPR/Cas9 system has ushered in a new era of targeted genetic manipulations. Here, we report the use of CRISPR/Cas9 to induce double-stranded breaks in the genome of the sea squirt Ciona intestinalis. We use electroporation to deliver CRISPR/Cas9 components for tissue-specific disruption of the Ebf (Collier/Olf/EBF) gene in hundreds of synchronized Ciona embryos. Phenotyping of transfected embryos in the 'F0' generation revealed that endogenous Ebf function is required for specification of Islet-expressing motor ganglion neurons and atrial siphon muscles. We demonstrate that CRISPR/Cas9 is sufficiently effective and specific to generate large numbers of embryos carrying mutations in a targeted gene of interest, which should allow for rapid screening of gene function in Ciona.
authors.affiliation: Center for Developmental Genetics, Department of Biology, New York University, New York, NY 10003, USA albertostolfi@gmail.com lc121@nyu.edu.
authors.forename: Alberto
authors.initials: A
authors.lastname: Stolfi
authors.affiliation: Center for Developmental Genetics, Department of Biology, New York University, New York, NY 10003, USA.
authors.forename: Shashank
authors.initials: S
authors.lastname: Gandhi
authors.affiliation: Center for Developmental Genetics, Department of Biology, New York University, New York, NY 10003, USA.
authors.forename: Farhana
authors.initials: F
authors.lastname: Salek
authors.affiliation: Center for Developmental Genetics, Department of Biology, New York University, New York, NY 10003, USA albertostolfi@gmail.com lc121@nyu.edu.
authors.forename: Lionel
authors.initials: L
authors.lastname: Christiaen
mesh_terms: D000818:Animals; D002938:Ciona intestinalis / Q000196:embryology* / Q000235:genetics / Q000378:metabolism*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics / Q000502:physiology*; D016678:Genome / Q000235:genetics*
keywords: Ascidians; CRISPR/Cas9; Ciona; Ebf; Genome editing; Islet