scholarly journals Highly efficient transgene‐free targeted mutagenesis and single‐stranded oligodeoxynucleotide‐mediated precise knock‐in in the industrial microalga Euglena gracilis using Cas9 ribonucleoproteins

2019 ◽  
Vol 17 (11) ◽  
pp. 2032-2034 ◽  
Author(s):  
Toshihisa Nomura ◽  
Komaki Inoue ◽  
Yukiko Uehara‐Yamaguchi ◽  
Koji Yamada ◽  
Osamu Iwata ◽  
...  
Keyword(s):  
Euglena Gracilis ◽  
Targeted Mutagenesis ◽  
Highly Efficient

scholarly journals Highly efficient targeted mutagenesis in one-cell mouse embryos mediated by the TALEN and CRISPR/Cas systems

2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Akihiro Yasue ◽  
Silvia Naomi Mitsui ◽  
Takahito Watanabe ◽  
Tetsushi Sakuma ◽  
Seiichi Oyadomari ◽  
...  
Keyword(s):  
Mouse Embryos ◽  
Targeted Mutagenesis ◽  
Highly Efficient

scholarly journals Highly Efficient Targeted Gene Editing in Upland Cotton Using the CRISPR/Cas9 System

2018 ◽  
Vol 19 (10) ◽  
pp. 3000 ◽  
Author(s):  
Shouhong Zhu ◽  
Xiuli Yu ◽  
Yanjun Li ◽  
Yuqiang Sun ◽  
Qianhao Zhu ◽  
...  
Keyword(s):  
Gene Editing ◽  
Crop Improvement ◽  
Targeted Mutagenesis ◽  
Cotton Fibers ◽  
Efficient Tool ◽  
Gene Encoding ◽  
Editing Event ◽  
Highly Efficient ◽  
Target Sites ◽  
Targeted Gene Editing

The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) gene editing system has been shown to be able to induce highly efficient mutagenesis in the targeted DNA of many plants, including cotton, and has become an important tool for investigation of gene function and crop improvement. Here, we developed a simple and easy to operate CRISPR/Cas9 system and demonstrated its high editing efficiency in cotton by targeting-ALARP, a gene encoding alanine-rich protein that is preferentially expressed in cotton fibers. Based on sequence analysis of the target site in the 10 transgenic cottons containing CRISPR/Cas9, we found that the mutation frequencies of GhALARP-A and GhALARP-D target sites were 71.4–100% and 92.9–100%, respectively. The most common editing event was deletion, but deletion together with large insertion was also observed. Mosaic mutation editing events were detected in most transgenic plants. No off-target mutation event was detected in any the 15 predicted sites analyzed. This study provided mutants for further study of the function of GhALARP in cotton fiber development. Our results further demonstrated the feasibility of use of CRISPR/Cas9 as a targeted mutagenesis tool in cotton, and provided an efficient tool for targeted mutagenesis and functional genomics in cotton.

scholarly journals Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis

2020 ◽  
Vol 1 (1) ◽  
pp. 100023
Author(s):  
Toshihisa Nomura ◽  
Mizuki Yoshikawa ◽  
Kengo Suzuki ◽  
Keiichi Mochida
Keyword(s):  
Genome Editing ◽  
Euglena Gracilis ◽  
Highly Efficient

scholarly journals Highly efficient targeted mutagenesis in axolotl using Cas9 RNA-guided nuclease

Development ◽  
2014 ◽  
Vol 141 (10) ◽  
pp. 2165-2171 ◽  
Author(s):  
G. P. Flowers ◽  
A. T. Timberlake ◽  
K. C. Mclean ◽  
J. R. Monaghan ◽  
C. M. Crews
Keyword(s):  
Targeted Mutagenesis ◽  
Highly Efficient

scholarly journals Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 System

Cell Reports ◽  
2014 ◽  
Vol 6 (6) ◽  
pp. 1178-1179 ◽  
Author(s):  
Andrew R. Bassett ◽  
Charlotte Tibbit ◽  
Chris P. Ponting ◽  
Ji-Long Liu
Keyword(s):  
Targeted Mutagenesis ◽  
Highly Efficient

Highly efficient multiplex targeted mutagenesis and genomic structure variation in Bombyx mori cells using CRISPR/Cas9

2014 ◽  
Vol 49 ◽  
pp. 35-42 ◽  
Author(s):  
Yuanyuan Liu ◽  
Sanyuan Ma ◽  
Xiaogang Wang ◽  
Jiasong Chang ◽  
Jie Gao ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Genomic Structure ◽  
Targeted Mutagenesis ◽  
Structure Variation ◽  
Highly Efficient

scholarly journals Highly Efficient Targeted Mutagenesis in Mice Using TALENs

Genetics ◽  
2013 ◽  
Vol 195 (3) ◽  
pp. 703-713 ◽  
Author(s):  
Sudeepta Kumar Panda ◽  
Benedikt Wefers ◽  
Oskar Ortiz ◽  
Thomas Floss ◽  
Bettina Schmid ◽  
...  
Keyword(s):  
Targeted Mutagenesis ◽  
Highly Efficient

Soluble and Particulate Porphobilinogen-Deaminase from Dark-Grown Euglena gracilis

1989 ◽  
Vol 44 (7-8) ◽  
pp. 578-580 ◽  
Author(s):  
M. V. Rossetti ◽  
A. A. Juknat ◽  
A. M. del C. Batlle
Keyword(s):  
Efficient Method ◽  
Euglena Gracilis ◽  
Equilibrium Mixture ◽  
Active Species ◽  
Porphobilinogen Deaminase ◽  
Chromatographic Behaviour ◽  
Highly Efficient ◽  
Chaotropic Agent ◽  
Molecular Masses ◽  
Membrane Bound

A highly efficient method employing NaSCN as a chaotropic agent was used to dissociate the membrane-bound porphobilinogen-deaminase. The same sequence of steps was applied for purifying both soluble and membrane dissociated porphobilinogen-deam inase. The chromatographic behaviour of both proteins was quite similar. Euglena gracilis deam inase appears to exist in an equilibrium mixture of two active species of relative molecular masses of 40000 and 20000.

scholarly journals Optimization of multiplexed CRISPR/Cas9 system for highly efficient genome editing in Setaria viridis

2020 ◽  
Author(s):  
Trevor Weiss ◽  
Chunfang Wang ◽  
Xiaojun Kang ◽  
Hui Zhao ◽  
Maria Elena Gamo ◽  
...  
Keyword(s):  
Genome Editing ◽  
Transformation Method ◽  
Targeted Mutagenesis ◽  
Setaria Viridis ◽  
Photosynthetic Pathway ◽  
Repair Pathway ◽  
End Joining ◽  
Highly Efficient ◽  
Homozygous Mutations ◽  
High Throughput Manner

AbstractIn recent years, Setaria viridis has been developed as a model plant to better understand the C4 photosynthetic pathway in major crops. With the increasing availability of genomic resources for S. viridis research, highly efficient genome editing technologies are needed to create genetic variation resources for functional genomics. Here, we developed a protoplast assay to rapidly optimize the multiplexed CRISPR/Cas9 system in S. viridis. Targeted mutagenesis efficiency was further improved by an average of 1.4-fold with the exonuclease, Trex2. Distinctive mutation profiles were found in the Cas9_Trex2 samples with 94% of deletions larger than 10bp, and less than 1% of mutations being insertions. Further analyses indicated that 52.2% of deletions induced by Cas9_Trex2, as opposed to 3.5% by Cas9 alone, were repaired through microhomology-mediated end joining (MMEJ) rather than the canonical NHEJ DNA repair pathway. Combined with the robust agrobacterium-mediated transformation method with more than 90% efficiency, the multiplex CRISPR/Cas9_Trex2 system was demonstrated to induce targeted mutations in two tightly linked genes, svDrm1a and svDrm1b, at the frequency ranging from 73% to 100% in T0 plants. These mutations were transmitted to at least 60% of the transgene-free T1 plants with 33% of them containing bi-allelic or homozygous mutations in both genes. This highly efficient multiplex CRISPR/Cas9_Trex2 system makes it possible to create a large mutant resource for S. viridis in a rapid and high throughput manner, and has the potential to be widely applicable in achieving more predictable MMEJ-mediated mutations in many plant species.

Sign in / Sign up

Export Citation Format

Share Document