ヨシダ ケイタ   YOSHIDA Keita
  吉田 慶太
   所属   医学部 医学科
   職種   助教
論文種別 原著
言語種別 英語
査読の有無 査読あり
表題 CRISPR/Cas9-mediated gene knockout in the ascidian Ciona intestinalis.
掲載誌名 正式名:Development, growth&differentiation
略  称:Dev Growth Differ
ISSNコード:1440-169X(Electronic)0012-1592(Linking)
掲載区分国外
巻・号・頁 56(7),499-510頁
著者・共著者 Sasaki Haruka†, Yoshida Keita, Hozumi Akiko, Sasakura Yasunori*
発行年月 2014/09
概要 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
文献番号 25212715