We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha.
- Authors
Sugano, Shigeo S.; Nishihama, Ryuichi; Shirakawa, Makoto; Takagi, Junpei; Matsuda, Yoriko; Ishida, Sakiko; Shimada, Tomoo; Hara-Nishimura, Ikuko; Osakabe, Keishi; Kohchi, Takayuki
- Abstract
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.
- Subjects
MARCHANTIA polymorpha; CRISPRS; GENOME editing; PLANT genomes; ARABIDOPSIS thaliana; NUCLEOTIDE sequence
- Publication
PLoS ONE, 2018, Vol 13, Issue 10, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0205117