期刊
DEVELOPMENT
卷 141, 期 21, 页码 4115-4120出版社
COMPANY BIOLOGISTS LTD
DOI: 10.1242/dev.114488
关键词
Ascidians; CRISPR/Cas9; Ciona; Ebf; Genome editing; Islet
资金
- American Heart Association [10SDG4310061]
- National Heart, Lung, and Blood Institute [5R01HL108643]
- New York Cardiac Center and New York University College of Arts and Sciences
- National Science Foundation Postdoctoral Research Fellowship in Biology [NSF-1161835]
- Div Of Biological Infrastructure
- Direct For Biological Sciences [1161835] Funding Source: National Science Foundation
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.
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