YOSHIDA Keita    Department   School of Medicine, School of Medicine    Position   Assistant Professor
Article types	Original article
Language	English
Peer review	Peer reviewed
Title	CRISPR/Cas9-mediated gene knockout in the ascidian Ciona intestinalis.
Journal	Formal name：Development, growth&differentiation Abbreviation：Dev Growth Differ ISSN code：1440-169X(Electronic)0012-1592(Linking)
Domestic / Foregin	Foregin
Volume, Issue, Page	56(7),pp.499-510
Author and coauthor	Sasaki Haruka†, Yoshida Keita, Hozumi Akiko, Sasakura Yasunori*
Authorship	2nd author
Publication date	2014/09
Summary	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.
DOI	10.1111/dgd.12149
Document No.	25212715