CRISPR/Cas9-mediated gene knockout in the ascidian Ciona intestinalis

CRISPR/Cas9-mediated gene knockout in the ascidian Ciona intestinalis. Sasaki, H., Yoshida, K., Hozumi, A., & Sasakura, Y. Development, Growth & Differentiation, 56(7):499–510, September, 2014.

Paper doi abstract bibtex

Knockout of genes with CRISPR/Cas9 is a newly emerged approach to investigate functions of genes in various organisms. We demonstrate that CRISPR/Cas9 can mutate endogenous genes of the ascidian Ciona intestinalis, a splendid model for elucidating molecular mechanisms for constructing the chordate body plan. Short guide RNA (sgRNA) and Cas9 mRNA, when they are expressed in Ciona embryos by means of microinjection or electroporation of their expression vectors, introduced mutations in the target genes. The specificity of target choice by sgRNA is relatively high compared to the reports from some other organisms, and a single nucleotide mutation at the sgRNA dramatically reduced mutation efficiency at the on-target site. CRISPR/Cas9-mediated mutagenesis will be a powerful method to study gene functions in Ciona along with another genome editing approach using TALE nucleases.

@article{sasaki_crispr/cas9-mediated_2014,
	title = {{CRISPR}/{Cas9}-mediated gene knockout in the ascidian {Ciona} intestinalis},
	volume = {56},
	copyright = {© 2014 The Authors Development, Growth \& Differentiation © 2014 Japanese Society of Developmental Biologists, This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.},
	issn = {1440-169X},
	url = {http://onlinelibrary.wiley.com/doi/10.1111/dgd.12149/abstract},
	doi = {10.1111/dgd.12149},
	abstract = {Knockout of genes with CRISPR/Cas9 is a newly emerged approach to investigate functions of genes in various organisms. We demonstrate that CRISPR/Cas9 can mutate endogenous genes of the ascidian Ciona intestinalis, a splendid model for elucidating molecular mechanisms for constructing the chordate body plan. Short guide RNA (sgRNA) and Cas9 mRNA, when they are expressed in Ciona embryos by means of microinjection or electroporation of their expression vectors, introduced mutations in the target genes. The specificity of target choice by sgRNA is relatively high compared to the reports from some other organisms, and a single nucleotide mutation at the sgRNA dramatically reduced mutation efficiency at the on-target site. CRISPR/Cas9-mediated mutagenesis will be a powerful method to study gene functions in Ciona along with another genome editing approach using TALE nucleases.},
	language = {en},
	number = {7},
	urldate = {2015-03-03},
	journal = {Development, Growth \& Differentiation},
	author = {Sasaki, Haruka and Yoshida, Keita and Hozumi, Akiko and Sasakura, Yasunori},
	month = sep,
	year = {2014},
	keywords = {Sasakura Y},
	pages = {499--510}
}

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