RRC ID 31337
Author Sasaki H, Yoshida K, Hozumi A, Sasakura Y.
Title CRISPR/Cas9-mediated gene knockout in the ascidian Ciona intestinalis.
Journal Dev. Growth Differ.
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.
Volume 56(7)
Pages 499-510
Published 2014-9
DOI 10.1111/dgd.12149
PMID 25212715
PMC PMC4231237
MeSH Animals CRISPR-Cas Systems / genetics CRISPR-Cas Systems / physiology* Ciona intestinalis / genetics* DNA Primers / genetics Electroporation Gene Knockout Techniques / methods* Genes, Homeobox / genetics Genetic Vectors / genetics Genomics / methods Mutagenesis / genetics* Mutation Rate Polymerase Chain Reaction
IF 1.496
Times Cited 18
C.intestinalis / (O.japonicus) Wild C. int