Germ cell mutations of the ascidian Ciona intestinalis with TALE nucleases
Abstract
Summary: Targeted mutagenesis of genes‐of‐interest, or gene‐knockout, is a powerful method to address the functions of genes. Engineered nucleases have enabled this approach in various organisms because of their ease of use. The ascidian Ciona intestinalis is an excellent organism to analyze gene functions by means of genetic technologies. In our previous study, we reported mutagenesis of Ciona somatic cells with TALE nucleases (TALENs) by electroporating expression constructs. In this study, we report germ cell mutagenesis of Ciona by microinjecting mRNAs encoding TALENs. TALEN mRNAs introduced mutations to target genes in both somatic and germ cells. TALEN‐mediated mutations in the germ cell genome were inherited by the next generation. We conclude that knockout lines of Ciona that have disrupted target genes can be established through TALEN‐mediated germ cell mutagenesis. genesis 52:431–439, 2014.Keywords
This publication has 25 references indexed in Scilit:
- Efficient Targeted Mutagenesis in Medaka Using Custom-Designed Transcription Activator-Like Effector NucleasesGenetics, 2013
- Efficient targeted gene disruption in Xenopus embryos using engineered transcription activator-like effector nucleases (TALENs)Proceedings of the National Academy of Sciences, 2012
- Efficient targeted mutagenesis of the chordate Ciona intestinalis genome with zinc‐finger nucleasesDevelopment, Growth & Differentiation, 2012
- Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targetingNucleic Acids Research, 2011
- Unfolding a chordate developmental program, one cell at a time: Invariant cell lineages, short-range inductions and evolutionary plasticity in ascidiansDevelopmental Biology, 2009
- Targeted gene inactivation in zebrafish using engineered zinc-finger nucleasesNature Biotechnology, 2008
- Efficient target-selected mutagenesis in Caenorhabditis elegans: Toward a knockout for every geneGenome Research, 2007
- Notch signaling is involved in nervous system formation in ascidian embryosWilhelm Roux' Archiv für Entwicklungsmechanik der Organismen, 2002
- Introduction and Expression of Recombinant Genes in Ascidian EmbryosDevelopment, Growth & Differentiation, 1992
- Altering the Genome by Homologous RecombinationScience, 1989