ID 112916
Title Alternative
Efficient genome editing and its application to conditional genetic analysis in M. polymorpha
Author
Sugano, Shigeo S. Ritsumeikan University|JST
Nishihama, Ryuichi Kyoto University
Shirakawa, Makoto Kyoto University
Takagi, Junpei Kyoto University
Matsuda, Yoriko Kyoto University
Ishida, Sakiko Kyoto University
Shimada, Tomoo Kyoto University
Hara-Nishimura, Ikuko Kyoto University
Kohchi, Takayuki Kyoto University
Content Type
Journal Article
Description
Marchantia polymorpha is one of the model species of basal land plants. Although CRISPR/ Cas9-based genome editing has already been demonstrated for this plant, the efficiency was too low to apply to functional analysis. In this study, we show the establishment of CRISPR/Cas9 genome editing vectors with high efficiency for both construction and genome editing. Codon optimization of Cas9 to Arabidopsis achieved over 70% genome editing efficiency at two loci tested. Systematic assessment revealed that guide sequences of 17 nt or shorter dramatically decreased this efficiency. We also demonstrated that a combinatorial use of this system and a floxed complementation construct enabled conditional analysis of a nearly essential gene. This study reports that simple, rapid, and efficient genome editing is feasible with the series of developed vectors.
Journal Title
PLOS ONE
ISSN
19326203
Publisher
PLOS
Volume
13
Issue
10
Start Page
e0205117
Published Date
2018-10-31
Rights
© 2018 Sugano et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
Publisher
departments
Bioscience and Bioindustry